Guidelines for the establishment and functioning of Animal Ethics Commitees (Institutional Animal Care and Use Committees) in Africa.

Animals are used for scientific purposes across Africa to benefit humans, animals or the environment. Nonetheless, ethical and regulatory oversight remains limited in many parts of the continent. To strengthen this governance framework, the Pan-African Network for Laboratory Animal Science and Ethics brought together experts from 12 African countries to create an Africa-centric practical guide to facilitate the establishment and appropriate functioning of Institutional Animal Ethics Committees across Africa. The Guidelines are based on universal principles for the care and use of sentient animals for scientific purposes, with consideration of the cultural, religious, political and socio-economic diversity in Africa. They focus on 11 key elements, including responsibilities of institutions and of the Institutional Official; composition of the Committee; its responsibilities, functioning and authority; ethical application and review processes; oversight and monitoring of animal care and use and of training and competence; quality assurance; and the roles of other responsible parties. The intent is for African institutions to adopt and adapt the guidelines, aligning with existing national legislation and standards where relevant, thus ensuring incorporation into practice. More broadly, the Guidelines form an essential component of the growing discourse in Africa regarding moral considerations of, and appropriate standards for, the care and use of animals for scientific purposes. The increased establishment of appropriately functioning animal ethics committees and robust ethical review procedures across Africa will enhance research quality and culture, strengthen societal awareness of animals as sentient beings, improve animal well-being, bolster standards of animal care and use, and contribute to sustainable socio-economic development.

Seasonal variations of testis anatomy and of G-coupled oestrogen receptor 1 expression in Gerbillus gerbillus.

The gerbil, Gerbillus gerbillus, a nocturnal desert rodent of northern Africa, exhibits a seasonal reproductive cycle with marked anatomical and behavioural shifts between breeding season and resting season. The aim of this study is to investigate key elements involved in these seasonal changes, specifically in males: the histology of the testis as well as the expression of the G-protein-coupled oestrogen receptor 1 (GPER1) in the testis. During the breeding season, the seminiferous tubules were full of spermatozoa, and their epithelium contained germinal cells embedded in Sertoli cells. Amidst tubules, well-developed Leydig cells were observed around blood vessels, with peritubular myoid cells providing structural and dynamic support to the tubules. GPER1 was largely expressed throughout the testis. Notably, Leydig cells, spermatogonia and spermatocytes showed strong immunohistochemical signals. Sertoli cells, spermatozoa and peritubular myoid cells were moderately stained. During the resting season, spermatogenesis was blocked at the spermatocyte stage, spermatids and spermatozoa were absent and the interstitial space was reduced. The weight of the testis decreased significantly. At this stage, GPER1 was found in Leydig cells, spermatocytes and peritubular myoid cells. Sertoli cells and spermatogonia were not marked. Overall, the testis of the gerbil, Gerbillus gerbillus, has undergone noticeable histological, cellular and weight changes between seasons. In addition, the seasonal expression pattern of GPER1, with pronounced differences between resting season and breeding season, indicates that this receptor is involved in the regulation of the reproductive cycle.

Impact of the nutritional quality of barley on growth rate, biometric and biochemical parameters and plasma concentration of androgens during puberty in the Ouled-Djellal lambs.

Puberty is part of physiological processes including growth, adrenarche, menarche, energy balance and metabolism. This study describes the dynamic between both metabolic and reproductive statutes during pubertal growth in Saharan breed sheep. Once weaned (3 months age), two lots of lambs are made up and each one receive a barley supplementation ration of 250 vs 500 g/head/day in addition to season's diet. Biometric measurements and blood samples are collected once a month from 3 to 12 months of age in order to evaluate biochemical and sexual hormonal status. Results show a significant weight gain and growth level in the double dose lot. Changes in biochemical parameters are closely related with age at least for glycemia and total proteinemia. Androgenic profile shows individual fluctuations (0.02 to 3.47 µg /ml) due to age, season and feeding ratio. In accordance with our findings, the diet effect is clearly evidenced between the two batches, it's noted that plasma concentration of androgens is the lowest (<0.30 ng /ml) at 3 months and increases to 0.53 vs 0.76 ng /ml between 4 and 6 months confirming the pre-pubertal phase. Also, biometric and biochemical parameters are tightly correlates with plasma androgen changes, depending on whether the animal be pubescent or not. In conclusion, although interesting this study shows no early puberty onset in the barley supplemented lambs as was reported in other sheep breeds; nevertheless, the testis activity as well as the body fitness have clearly be enhance. The synergy between biochemical profiles and biometric measurements explain the metabolic function of testicular androgens at puberty.

Multifloral white honey outclasses manuka honey in methylglyoxal content: assessment of free and encapsulated methylglyoxal and anti-microbial peptides in liposomal formulation against toxigenic potential of Bacillus subtilis Subsp spizizenii strain.

The therapeutic virtues of honey no longer need to be proven. Honey, which is rich in nutrients, is an excellent nutritional food because of its many properties; however, honey has been diverted from this primary function and used in clinical research. Evidence has shown that honey still possesses unknown properties and some of these aspects have never been addressed. In this work, two bioactive compounds found in honey (methylglyoxal and antimicrobial peptides) were evaluated for their anti-Bacillus subtilis activity with particular attention to their dilution factor. Although this bacterial strain does not possess an indigenous virulence factor gene, it becomes virulent by transferring plasmids with B. thuringiensis or expression of toxins from Bordetella pertussis. As is known, methylglyoxal is a toxic electrophile present in many eukaryotic and prokaryotic cells, which is generated by enzymatic and non-enzymatic reactions. Its overexpression successfully kills bacteria by inducing membrane disruption. Also, AMPs show potent inhibitory action against Gram-positive bacteria. Because of the lack of information concerning the main ingredients of honey, the microencapsulation process was used. Both methylglyoxal (MGO) and peptide-loaded liposomes were synthesized, characterized and compared to their free forms. The liposomal formulations contained a mixture of eggPC, cholesterol, and octadecylamine and their particle sizes were measured and their encapsulation efficacy calculated. The results revealed that Algerian multifloral white honey contained higher levels of MGO compared to manuka honey, which prevented bacterial growth and free MGO was relatively less effective. In fact, MGO killed BS in the loaded form with the same bacteriostatic and bactericidal index. However, the action of AMPs was different. Indeed, the investigation into the reactivity of MGO in the solvent indicated that regardless of the level of water added, honey is active at a fixed dilution. This data introduces the notion of dilution and abolishes the concept of concentration. Moreover, the synergistic antibacterial effect of the compounds in honey was diminished by the matrix effect. The degree of liposome-bacteria-fusion and the delay effect observed could be explain by both the composition and nature of the lipids used. Finally, this study reinforces the idea that under certain conditions, the metalloproteinases in honey produce AMPs.

Endocrine, energy, and lipid status during parturition and early lactation in indigenous goats native to the Algerian Sahara.

BACKGROUND AND AIM: Goats are widely distributed in southwest Algeria. The Saharan goat is perfectly adapted to the harsh conditions of arid areas, and it is characterized by resistance to long photoperiod and reduced metabolic needs, allowing the survival of its offspring by maintaining lactation. Several studies have demonstrated that parturition and lactation are critical periods that induce hormone, energy, and lipid status changes in mammals. However, the relationship between the blood biochemical parameters of parturition control and lactation functions in the Algerian Saharan goat has not been thoroughly documented. Therefore, this study assesses hormone and metabolite levels during parturition and early lactation in Saharan goats reared in arid areas. MATERIALS AND METHODS: Experiments were performed on 14 multiparous female goats, and blood samples were collected during parturition, 4 days postpartum (D1PP-D4PP), and during the first 12 weeks of lactation (W1-W12) to analyze prolactin, cortisol, glucose (GLU), total proteins (TP), cholesterol (CHO), triglycerides (TGs), total lipids (TL), low-density lipoproteins (LDLs), high-density lipoproteins (HDLs), and very LDLs (VLDLs). RESULTS: Statistical data analysis revealed a significant (p<0.05) increase in plasma prolactin concentrations at W1 after parturition, reaching maximum values at W3 and W9, and remained high until W12 of lactation. Plasma cortisol levels were high at parturition, reaching two peaks at W3 and W9, and then decreased at W5, W7, and W12 of lactation. No significant changes were found in serum GLU levels during the first 7 weeks of lactation compared with parturition day; then, the levels became significantly (p<0.05) lower at W8, W11, and W12 of lactation. Plasma TP increased significantly (p<0.05) at D3PP, W1, and W4, then decreased significantly (p<0.05) at W8. In addition, this decrease coincided with that of GLU production. Serum CHO, TGs, TL, LDLs, and VLDLs, were low at parturition and high at D4PP and during the first 3 months of lactation. Furthermore, HDL levels were low at D3PP, 1st, and 3rd months and high at the 2nd month of lactation. CONCLUSION: This study emphasized the impact of parturition and the 1st weeks of lactation on endocrine and metabolic changes in indigenous goats living in the Algerian Sahara Desert. These results can be used to monitor and improve farming management and understand physiological adaptive strategies, mainly lactation function sustainability, of this goat living in marginal zones.

Molecular docking and dynamic studies of a potential therapeutic target inhibiting glyoxalase system: Metabolic action of the 3, 3 '- [3- (5-chloro-2-hydroxyphenyl) -3-oxopropane-1, 1-diyl] - Bis-4-hydroxycoumarin leads overexpression of the intracellular level of methylglyoxal and induction of a pro-apoptotic phenomenon in a hepatocellular carcinoma model.

Methylglyoxal is a dicarbonyl compound recruited as a potential cytotoxic marker, initially presents in cells and considered as a metabolite of the glycolytic pathway. Our aim is to demonstrate the inhibitory effect of 3, 3'-[3-(5-chloro-2-hydroxyphenyl)-3-oxopropane-1, 1-diyl] Bis (4-hydroxycoumarin) on the glyoxalase system, and indirectly its anticancer activity. The docking of OT-55 was conducted by using Flexible docking protocol, ChiFlex and libdock tools inside the active site of Glo-I indicated that both hydrogen bonding and hydrophobic interactions contributed significantly in establishing potent binding with the active site which is selected as a strong inhibitor with high scoring values and maximum Gibbs free energy. Coumarin-liposome formulation was characterized and evaluated in vivo against chemically induced hepatocarcinoma in Wistar rats. After Diethylnitrosamine (DEN) induction, microscopic assessment was realized; precancerous lesions were developed showing an increase of both tumor-associated lymphocyte and multiple tumor acini supported by the blood investigation. Our finding also suggested a preferential uptake of liposomes respectively in liver, kidney, lung, brain and spleen in the DEN-treated animals. OT-55 has also been shown to inhibit the activity of Glo-I in vitro as well as in DEN-treated rats. An abnormal high level of MGO of up to 50% was recorded followed by a reduction in glucose consumption and lactate dehydrogenase production validated in the positive control. MGO generates apoptosis as depicted by focal hepatic lesions. Also, no deleterious effects in the control group were observed after testing our coumarin but rather a vascular reorganization leading to nodular regenerative hyperplasia. Involved in the detoxification process, liver GSH is restored in intoxicated rats, while no changes are seen between controls. At the endothelial cell, OT-55 appears to modulate the release of NO only in the DEN-treated group. OT-55 would behave both as an anticancer agent but also as an angiogenic factor regarding results obtained.

Assessment of Testicular Lhcgr mRNA Expression Correlated with Testis and Seminal Vesicle Activities in the Libyan jird (Meriones libycus, Rodentia: Muridae) during Breeding Season Compared with Nonbreeding Season.

The Libyan jird (Meriones libycus, 1823) is a wild desert rodent that is a seasonal breeder species adapted to breed when the environmental conditions can satisfy the energy and hydrous requirements of pregnant and nursing females to ensure that births occur at the most favorable time of the year. We assessed gene expression of testicular luteinizing hormone receptor (Lhcgr) correlated to testis activity. The expression of Lhcgr was evaluated using quantitative Real Time-Polymerase Chain Reaction (qRT-PCR and the testis activity by a histological method in adult male Libyan jirds during the nonbreeding and breeding seasons. Our results showed that Lhcgr mRNA expression increased in autumn during the nonbreeding season and decreased in spring during the breeding season. This expression varied in contrast to testicular structure or function and plasma testosterone levels. These results help to elucidate this desert rodent's seasonal sexual activity, which is correlated with central regulation.

Development of analytical methods GC-MS vs LC-UV for the serum monitoring of an inflammatory glycotoxin (methylglyoxal): A new biomarker of bovine hepatobiliary distomatosis.

Two analytical methods; high performance liquid chromatography and gas chromatography were used to determine the content of 2-methylquinoxaline, a methylglyoxal-derived agent in sera from cattle with fascioliasis. Methylglyoxal is a highly mutagenic and cytotoxic reactive dicarbonyl compound formed by non-enzymatic fragmentation of triose phosphate GAP and DHAP during glycolysis which regularly contributes to repositioning the energetic balance between physiological and pathological situations. The aim of this study was to propose the MGO as a new biomarker in the bovine fasciolosis. Strongly infected animals showed a correlation between the relatively high levels of Fasciola hepatica anti-f2 antibody and methylglyoxal compared to unharmed animals. Also, an acute hyperglycemia was recorded and closely related to hepatic parenchyma hyperplasia, inflammation, bile ducts obstruction and scléro-fibrous foci formation.Unlike HPLC, which has shown analytical flaws and irregularities, GC-MS remains an excellent diagnostic tool for detecting and quantifying methylglyoxal in biological fluids. The developed method has been validated under FDA guidelines. A full scan-range was set from m/z 39 to 144/999 and the molecular weight of the 2-methylquinoxaline was identified according to NIST Database and ES. Methylglyoxal was the only analyte successfully quantified in a relatively short run time. It was linear over a concentration range of 0.057-5.7  µg.ml-1with mean recoveries and RSD of 118% and 3.63% respectively. The intra and inter-day assays were satisfying and not exceed 3.00%. Results reflect the degree of precision of our method and indicate that MGO was an important contributor to understand the hepatic failure independently of other serum markers.

Seasonal changes in plasma testosterone and cortisol suggest an androgen mediated regulation of the pituitary adrenal axis in the Tarabul's gerbil Gerbillus tarabuli (Thomas, 1902).

In the desert gerbil Gerbillus tarabuli (Thomas, 1902), cortisol is the main glucocorticosteroid produced by the adrenal glands. Plasma cortisol concentrations show highest values when testosterone is reduced and lowest values during the breeding season which occurs from early winter to late spring. In order to specify the implication of testicular androgens in these corticosteroid seasonal variations we investigated the effects induced by gonadectomy performed during the breeding season on the pituitary adrenal axis. The animals collected in winter were assessed into three groups: sham-operated (Controls; n=13), gonadectomised (GDX; n=13) and testosterone replaced gonadectomised (GDX+T; n=13). Physiological replacement of testosterone enanthate (75µg/100gb.w./twice daily) was applied during one week, while GDX group received the vehicle (40µL sesame oil) alone. The right adrenal glands removed from euthanized animals were fixed for histomorphometry and androgen receptors (ARs) immunohistochemistry and the left ones were frozen with plasma samples until hormonal assays. Gonadectomy induces the enlargement of the adrenal cortex essentially due to that of zonae fasciculata (ZF) and reticularis (ZR) and perimedullary connective tissue which is abundant in the gerbil adrenals. The ARs immunostaining present at both cytoplasmic and nucleus level, is enhanced intensely in the ZR and moderately in the ZF and zona glomerulosa (ZG) cells. GDX group shows reduced plasma ACTH concentration (p=0.0126) by 61% despite the increase in cortisol concentration occurring both in plasma (+216%; p=0.0436) and adrenal tissue (+117%; p=0.0348). Plasma aldosterone is also enhanced significantly (p=0.0147) by 189% but androstenedione synthesis increased in adrenal tissue (p=0.0459) by 65% instead a decrease at circulatory level (p=0.0355) by 58% due to lack of testicular origin. So, testosterone deprivation activates corticosteroidogenesis also evidenced by the adrenal structure changes and the gonadectomy-induced increase in the plasma cholesterol. All of the gonadectomy-induced responses are reversible after physiological testosterone replacement. We conclude that the assessment of circulating adrenocorticotropic hormone (ACTH) concentrations together with cortisol levels essentially, reflecting the hypothalamic-pituitary adrenal (HPA) axis feedback loop control during the annual endogenous changes of testosterone secretion, represents a well-adapted response of this desert species living in an extreme environment.

GPER1 in sand rat epididymis: Effects of seasonal variations, castration and efferent ducts ligation.

Estrogen plays a crucial role in regulating epididymal function and development. Estrogen signaling is mediated via two main receptors essentially involved in the genomic regulating pathway: ERα and ERß. Recent studies revealed the contribution of a novel estrogen receptor involved in the non-genomic pathway: GPER1. This receptor belongs to the family of seven-transmembrane G-protein-coupled receptors and it triggers rapid cellular responses. Immuno-histochemical studies and Western Blot analyses were performed to investigate the GPER1 expression in the caput and cauda epididymis of free-ranging fat sand rats (Psammomys obesus) captured during the breeding and resting seasons. We also investigated the effect of castration (C), castration followed by testosterone treatment (C+T), and ligation of the efferent ducts (L). During the breeding season, a marked positive GPER1 immunoreactivity was detected in the cytoplasm of principal cells and basal cells; this signal persisted during the resting season, attenuated however, meanwhile the clear cells were not immuno-reactive. In C animals, the immuno-histochemical staining underwent nuclear translocation. In C+T animals, this response became nuclear and cytoplasmic. In the L group, the expression of the GPER1 was mainly located in the cytoplasm of principal cells and in the nuclei of basal cells; the sperm was also immune-positive in the cauda epididymis. Western blot analysis showed that GPER1 has a molecular weight of 55kDa in the caput and cauda epididymis during the breeding season, and it persisted during the resting season in the caput epididymis with a decrease in the cauda epididymis. These results suggest that GPER1 mediate a specific cellular estrogen signaling with marked differences between the breeding and resting seasons. Experimental groups suggest that testosterone is involved in the regulation of the expression of GPER1, in addition to other estrogen signalization pathways.

Seasonal variations of aromatase and estrogen receptors expression in the testis of free-ranging sand rats.

An increasing number of studies revealed the importance of estrogen in male reproduction. However, most research was conducted in laboratory rodents subjected to standardized environmental conditions. Therefore, seasonal regulations of estrogen pathways remain poorly understood under natural conditions. Using immunohistochemistry, the expression of several molecules involved in the functioning of testis (i.e. 17-ß estradiol [E2], P450 aromatase, estrogen receptors ESR1, ESR2, and GPER1 [also known as GPR30]) were investigated in free-ranging fat sand rats, Psammomys obesus, during the breeding and resting seasons. Leydig cells showed a strong immunoreactivity for aromatase in the testis sampled during the breeding season only; however, E2, ESR1, ESR2 and GPER1 were present during both seasons. Sertoli cells showed a positive signal for E2 and ESR2 during the breeding season; though, all molecules, except GPER1, were present during the resting season. Spermatogonia were reactive for E2, ESR2 and GPER1 during the breeding season and for ESR1 and GPER1 during the resting season. During both seasons, spermatocytes-I presented a moderate reactivity for E2, ESR1, ESR2 and a strong reactivity for GPER1; aromatase was detected during the resting season only. Spermatids and spermatozoa were present exclusively during breeding season and were reactive for all molecules; except round spermatids that were negative for aromatase. The functioning of the testis depends on finely tuned stimulation and inhibition systems. Our results suggest that differential expression of aromatase, ESR1, ESR2, and GPER1 across cells types is involved in the seasonal activation/inactivation cycle of spermatogenesis in a free-ranging species.

Seasonal morphophysiological variations in the prostatic complex of the Tarabul's gerbil (Gerbillus tarabuli).

Gerbillus tarabuli is a nocturnal Saharan rodent which has an annual reproductive cycle characterized by the reproductive activity in spring and a long phase of sexual quiescence in other seasons. We describe the morphology and hormonal regulation of the prostatic complex of this rodent in the two periods, based on anatomical, histological, morphometric, and immunohistochemical analyses. The organisation of this prostatic complex is similar to that reported for Meriones unguiculatus, but different from the prostate of Psammomys obesus, the rat, and the mouse. In addition to the anterior lobes, ventral lobes, and dorsal lobes, the prostatic complex of Gerbillus tarabuli, also includes dorsolateral lobe. Each lobe is composed of a fibro-muscular stroma surrounding a glandular epithelium. Dorsolateral lobes are easily distinguishable by their big volume. The prostate grows and regresses cyclically throughout the year. During the resting season, ventral lobes and anterior lobes showed atrophy, with a significant decrease in both epithelial height and supranuclear area size, and a strong thickening of the fibro-muscular compartment. In dorsal lobes, the epithelial and stromal compartments atrophied and regenerated simultaneously, whereas in dorsolateral lobe the thickness of the epithelium, the supranuclear zone and the stroma increased during resting period. Furthermore, seasonal variations were observed in the distribution and expression of both androgen receptors, and estrogens receptors. Expression patterns of all receptors were lobe-specific. In conclusion, both androgens and estrogens are involved in the homeostasis and regulation of the prostate in Gerbillus tarabuli. Dorsolateral lobe seems to be controlled by a different mechanism than other lobes.

Analysis of the energetic metabolism in cyclic Bedouin goats (Capra hircus): Nychthemeral and seasonal variations of some haematochemical parameters in relation with body and ambient temperatures.

Several studies have examined changes in some haematochemical parameters as a function of the different physiological status (cyclic, pregnant and lactating) of goats, but no relevant literature has exhaustively investigated these variations from anestrous to estrous stages in cyclic goats. In this paper, we report nychthemeral and seasonal variations in ambient and body temperatures, and in some haematochemical parameters (glycemia, cholesterolemia, triglyceridemia, creatininemia and uremia) measured during summer, winter and spring, in seven (7) experimental cyclic female Bedouin goats (Capra hircus) living in the Béni-Abbès region (Algerian Sahara desert). Cosinor rhythmometry procedure was used to determine the rhythmic parameters of ambient temperature and haematochemical parameters. To determine the effect of time of day on the rhythmicity of the studied parameters, as well as their seasonality, repeated measure analysis of variance (ANOVA) was applied. The results showed that in spite of the nychthemeral profile presented by the ambient temperature for each season, the body temperature remained in a narrow range, thus indicating a successful thermoregulation. The rhythmometry analysis showed a circadian rhythmicity of ambient temperature and haematochemical parameters with diurnal acrophases. A statistically significant effect of the time of day was shown on all studied haematochemical parameters, except on creatininemia. It was also found that only uremia, cholesterolemia and triglyceridemia followed the seasonal sexual activity of the studied ruminant. This study demonstrated the good physiological adaptation developed by this breed in response to the harsh climatic conditions of its natural environment.

Androgen receptor-mediated regulation of adrenocortical activity in the sand rat, Psammomys obesus.

The wild sand rat, Psammomys obesus, displays seasonal variations in adrenocortical activity that parallel those of testicular activity, indicating functional cross-talk between the hypothalamo-pituitary-adrenal and hypothalamo-pituitary-gonadal axes. In the present study, we examined androgen receptor (AR)-mediated actions of testicular steroids in the regulation of adrenocortical function in the sand rat. Specifically, we examined the expression of AR in the adrenal cortex, as well as adrenal apoptosis in male sand rats that had been surgically castrated or castrated and supplemented with testosterone; biochemical indices of adrenocortical function and hormone profiles were also measured. Orchiectomy was followed by an increase in adrenocorticotropic hormone secretion from the anterior pituitary and subsequently, increased adrenocortical activity; the latter was evidenced by orchiectomy-induced increases in the adrenal content of cholesterol and lipids as well as adrenal hypertrophy (seen as an elevation of the RNA/DNA ratio). Further, androgen deprivation respectively up- and downregulated the incidence of apoptosis within the glucocorticoid-producing zona fasciculata and sex steroid-producing zona reticularis. Interestingly, orchiectomy resulted in increased expression of AR in the zona fasciculata. All of the orchiectomy-induced cellular and biochemical responses were reversible after testosterone substitution therapy. Together, these data suggest that adrenocortical activity in the sand rat is seasonally modulated by testicular androgens that act through AR located in the adrenal cortex itself.

Immunolocalization of estrogen and androgen receptors in the caput epididymidis of the fat sand rat (Psammomys obesus): Effects of seasonal variations, castration and efferent duct ligation.

The fat sand rat (Psammomys obesus) is a model to study seasonal reproductive cycle changes and several metabolic disorders. In order to show a possible involvement of estrogens in the male reproductive functions, the expression of estrogen receptors (ESR1 and ESR2) and androgen receptor (AR) were investigated in the caput epididymidis of fat sand rats during the breeding season, resting season, after castration, after castration followed by testosterone treatment, and after ligation of efferent ducts. In the breeding season, principal cells presented a strong immunostaining of AR in both nuclei and cytoplasm, a strong staining of ESR1, mainly in the apical zone, and a strong immunoexpression of ESR2, mainly in nuclei. In the resting season, a moderate immunostaining of AR in both cytoplasm and nuclei was observed. ESR1 staining showed a strong immunoreactivity in the nuclei. In contrast, the nuclei were negative for ESR2. After castration, a low and selective signal distribution was observed: the nuclei were moderately positive for AR and ESR2, and negative for ESR1. After castration and testosterone treatment, an androgen-dependence for AR and the restoration of ESR1 but not ESR2 immunoexpression were observed. After ligation of the efferent ducts, a considerable reduction of AR immunoreactivity was observed in contrast to ESR1 and ESR2, which gave a strong immunostaining signal. These results illustrate the complexity of the regulation of the androgen and estrogen receptor expression in the epididymis and argue for the coexistence of both androgenic and estrogenic pathways.

Transgelin: an androgen-dependent protein identified in the seminal vesicles of three Saharan rodents.

During the breeding season, a major androgen-dependent protein with an apparent molecular weight of 21 kDa was isolated and purified from the seminal vesicles of three Saharan rodents (MLVSP21 from Meriones libycus, MSVSP21 from Meriones shawi, and MCVSP21 from Meriones crassus). The 21-kDa protein was isolated and purified from soluble seminal vesicle proteins of homogenate by one-dimensional polyacrylamide gel electrophoresis (SDS-PAGE). Using polyclonal antibodies directed against POSVP21 (Psammomys obesus seminal vesicles protein of 21 kDa), a major androgen-dependent secretory protein from sand rat seminal vesicles, identified previously as transgelin, we showed an immunological homology with POSVP21 by immunoblotting. These three major androgen-dependent proteins with a same apparent molecular weight of 21 kDa designated as MLVSP21 (Meriones libycus seminal vesicles protein of 21 kDa), MSVSP21 (Meriones shawi seminal vesicles protein of 21 kDa), and MCVSP21 (Meriones crassus seminal vesicles protein of 21 kDa) were localized by immunohistochemistry and identified by applying a proteomic approach. Our results indicated that the isolated proteins MLSVP21, MSSVP21, and MCSVP21 seem to correspond to the same protein: the transgelin. So that transgelin can be used as a specific marker of these rodent physiological reproduction mechanisms.

Postnatal changes in testicular development and androgen receptors immunolocalization in D'Man ram lambs.

The purpose of this study was to characterize testicular development in D'Man ram lambs, focusing primarily on androgen receptors (ARs) immunolocalization in the adenohypophysis and testis that is not still known in the D'Man ram lamb. Lambs (n = 12) were divided into four groups (three lambs per group). Adenohypophysis and testis were fixed and paraffin embedded; cross-section (3 µm) were stained and evaluated with immunohistochemistry. Testis weight increased at a greater rate between two and five months after birth, which was associated with remarkable changes in testicular histology, including significant increases in the diameter of seminiferous tubules. Spermatogenesis started between three and five months after birth; lumen and elongated spermatids were observed for the first time in three and four months-old animals respectively. ARs detected with immunohistochemistry were located in the nuclei and cytoplasm of adenohypophysis cells, and only in nuclei of testis cells (Leydig, Sertoli, peritubular myoid and germ cells).

Identification, cloning and regulation of cDNA encoding aldo-keto reductase 1B7 in the adrenal gland of two Saharan rodents Meriones libycus (Libyan jird) and Gerbillus gerbillus (gerbil).

Aldo-Keto Reductase 1B7 (AKR1B7) is a mouse aldose reductase-like protein with two major sites of expression, the vas deferens and the adrenal cortex. In the adrenal cortex, Akr1b7 is an adrenocorticotropin (ACTH)-responsive-gene whose product scavenges harmful byproducts of steroidogenesis and limits stress response through the biosynthesis of prostaglandin F2α. The purpose of the present study was to explore the possible expression of AKR1B7 in the adrenal glands of two saharan rodents, Libyan jird and Lesser Egyptian gerbil. Western blot analyses demonstrated that a protein related to murine/rat AKR1B7 was highly expressed in adrenals and absent from vas deferens of both saharan species. Based on conserved sequences between mouse and rat, full length cDNA were cloned and sequenced in both species while hormonal regulation and tissue localization were explored in Libyan jird. Both cDNA encoded the expected 316 amino acids protein typical of AKR1B subfamily and contained the highly conserved catalytic tetrad consisting in Asp-44, Tyr-49, Lys-78 and His-111 residues. The deduced proteins shared higher identities with aldose reductase-like, i.e. AKR1B7 (86-94%), AKR1B8 and AKR1B10 (83-86%) than with aldose reductase group, i.e. AKR1B1 and AKR1B3 (70%). Phylogenetic analysis showed that the Libyan jird and gerbil enzymes were more closely related to murine and rat AKR1B7 than to the other AKR1B members. Northern blot analyses of total RNA from Libyan jird adrenals showed a single mRNA transcript of 1.4 kb whose expression was dependent on circulating ACTH levels. In conclusion, we demonstrate here that adrenal glands of Libyan jird and gerbil express both an ortholog of the murine/rat Akr1b7 gene and that ACTH-responsiveness is at least conserved in Libyan jird.

Morphometry and immunohistochemistry of follicles growth and steroidogenesis in saharian wild sand rat, Psammomys obesus, ovary.

The sand rat (Psammomys obesus) constitutes a model to study seasonal changes and several metabolic disorders. In order to perform breeding laboratory conditions, the reproductive function of this species living in North Occidental Algerian Sahara was studied. The aim of this work was to investigate the follicular growth changes and the steroidogenic associated aspects. The study was performed using morphometrical and immunohistochemical methods. From primordial to preantral states, the follicle diameter increased progressively from 17-20 mum to 192-225 mum. The preovulatory follicles reached about 500 mum in diameter. Immunoreactivity to progesterone, androstenediol and estradiol, varied in the different parts of the ovary and follicular cells. The progesterone antibody appeared clearly labelled in the theca interna of the growing follicle and increased in the granulosa; the androgen antibody was continuously weak and diffuses in all follicles; the estradiol labelling appeared weak and diffuse in preantral follicles then increased in antral follicles in both theca and granulosa or only in granulosa. In antral follicles, estradiol label was clearly localized in granulosa cells and totally devoid in theca cells. In Psammomys ovary, labels of hormone were diffuse or localized, weak or intense in the theca and or in the granulosa according to the follicle size.

Histological and immunohistological aspects of the ovarian cycle of the algerian wild sand rat, Psammomys obesus Cretzschmar, 1828.

The sand rat, Psammomys obesus, is largely used as a model for studying several metabolic disorders. In order to perform breeding laboratory conditions, the reproductive function of this species was investigated. Using histological and immunohistochemical techniques, several aspects of the ovaries were studied throughout the sexual cycle. During the ovarian cycle, the different stages of folliculogenesis, from primordial to Graafian follicle, have been shown; the differentiation of both granulosa and theca cells, the formation of the antrum, cumulus oophorus and corona radiate were described. Broken follicles and corpora lutea have been observed, confirming a spontaneous ovulation in isolated females. Steroid activities were analysed using immunohistochemical techniques. Estrogen, androgen and progesterone hormones were visualized in the different compartments of the ovary.