Epistatic Effects Between Amino Acid Insertions and Substitutions Mediate Toxin resistance of Vertebrate Na+,K+-ATPases.

The recurrent evolution of resistance to cardiotonic steroids (CTS) across diverse animals most frequently involves convergent amino acid substitutions in the H1-H2 extracellular loop of Na+,K+-ATPase (NKA). Previous work revealed that hystricognath rodents (e.g., chinchilla) and pterocliform birds (sandgrouse) have convergently evolved amino acid insertions in the H1-H2 loop, but their functional significance was not known. Using protein engineering, we show that these insertions have distinct effects on CTS resistance in homologs of each of the two species that strongly depend on intramolecular interactions with other residues. Removing the insertion in the chinchilla NKA unexpectedly increases CTS resistance and decreases NKA activity. In the sandgrouse NKA, the amino acid insertion and substitution Q111R both contribute to an augmented CTS resistance without compromising ATPase activity levels. Molecular docking simulations provide additional insight into the biophysical mechanisms responsible for the context-specific mutational effects on CTS insensitivity of the enzyme. Our results highlight the diversity of genetic substrates that underlie CTS insensitivity in vertebrate NKA and reveal how amino acid insertions can alter the phenotypic effects of point mutations at key sites in the same protein domain.

Colocalization pattern of cocaine- and amphetamine-regulated transcript peptide and parvalbumin immunoreactivity in the hippocampus proper of the chinchilla.

This study set out to investigate, for the first time, the distribution and colocalization pattern of cocaine-and amphetamine-regulated transcript (CART) and one of the calcium binding- -proteins: parvalbumin (PV) in the chinchilla's hippocampus proper (HP). HP, consisting of Ammon's horn (CA) and the dentate gyrus (DG), is an important component of the limbic system, involved in learning and memory processes. CA showed a higher immunoreactivity of CART (-IR) compared to DG. CART-IR neurons were mainly observed in the molecular layer of DG and in the pyramidal layer of CA. CART-IR fibers were present in the granular layer; in the hilus numerous mossy fibers were detected, while in the molecular layer CART-IR fibers were not found. In all CA fields (CA1-CA3), CART-IR fibers were only present in the lacuno- sum-molecular layer. Immunofluorescence with double- labeling showed that only CART-IR cells stained positive for PV, whereas in CART-IR fibers there was no PV-positive reaction. Our research supplements missing knowledge about the distribution and colocalization pattern of CART with PV in the chinchilla's hippocampus, and also provides a better understanding of the similarities and differences among individuals of the same species and also with other mammals.

Analysis of glycoconjugates and morphological characterization of the descending colon and rectum of the plains viscacha, Lagostomus maximus.

Herbivores exhibit specializations at the intestinal level that facilitate the bacterial fermentation. The available information on the digestive physiology of Lagostomus maximus makes this rodent an interesting model to evaluate morpho-functional adaptations to herbivory. The general objective of this work was centered on the study of the morphology and histochemistry of the descending colon and rectum of L. maximus. To do so, a comparative analysis of the morphology, ultrastructure and glycosylation pattern of both anatomical regions was carried out. Histochemical results revealed that in both sectors of the large intestine, there are goblet cells with different glycosylation pattern within a morphologically homogeneous cell population. The main difference between both intestinal segments lay in the fact that the most distal region of the large intestine showed a greater proportion of sialomucins, characterized by being slightly O-acetylated. Further specific differences were revealed by lectin histochemistry. These data allowed to perform a functional interpretation of the cell types and secreted substances, thus contributing to a better understanding of the role of mucins in the intestinal tract functioning.

Oviduct morphology and estrogen receptors ERα and ERß expression in captive Chinchilla lanigera (Hystricomorpha: Chinchillidae).

Chinchilla lanigera is a hystricomorph rodent from South America whose reproductive biology presents particular characteristics that distinguishes it from other Rodentia species, such as low reproductive rate, seasonal breeding pattern, and long estrous cycle. Nevertheless, reproductive features in female chinchillas are still poorly investigated, with a scarce knowledge concerning the estrous cycle and the histology of reproductive organs. In this study, we investigate the morphology, histomorphometry, secretory activity, and immunolocalization of estrogen receptors ERα and ERß in oviducts of nulliparous chinchillas, euthanized at fall season in Brazil. Follicular phase of estrous cycle of all studied animals was characterized by ovary and uterine morphology inspection, as well as vaginal cytology. Similar to other mammals, the oviduct wall of infundibulum, ampulla and isthmus was composed of mucosa, muscle, and serosa layers. Morphometric data of oviduct layers were used for identifying each oviduct segment. In the follicular phase, the oviduct was characterized by intense secretory activity, mainly in the ampulla, and expression of ERα and ERß throughout the oviduct epithelium. Both ERα and ERß were also detected in the connective tissue and smooth muscle cells. Our findings point out to the important role of estrogen in this female organ. Similar wide distribution of both ER proteins has been described for human Fallopian tube. Taken together, our data add to the understanding of the reproductive biology of female chinchillas, and may assist in the intensive breeding of this species and any eventual endeavor for conservation of chinchillas in the wild.

Immunocytochemical detection of calretinin in the claustrum and endopiriform nucleus of the chinchilla.

Calretinin (CR) as a buffer and sensor protein plays an important role in regulatory processes of Ca²âº and anty-apoptotic cellular protection. In the present study, immunohistochemical peroxidase-antiperoxidase (PAP) method was used in order to determine the numbers, morphology, morphometry and distribution pattern of CR in neurons of the chinchilla's claustrum (Cl) and endopiriform nucleus (EN). In Cl and EN the presence of several classes of neurons with different immunoreactivity to CR was found. In Cl, CR-immunoreactive (IR) neurons were predominantly found in close vicinity to insular border while CR-IR neurons were evenly scattered throughout EN. In general, immunoreaction to CR was observed in neuronal cytoplasm, nucleus and in fibres-like nerve extensions. Statistical analysis showed the differences between average large diameter as well as cross-sectional area of CR-IR neurons present in Cl and EN. It is suggested, that CR acting as a calcium binding protein may play a role in neuronal network. Further co-localization studies are necessary to fully elucidate neurophysiology and neuropathology of the chinchilla's Cl and EN neurons.

Seasonal evaluations of urinary androgen metabolites and semen quality in domestic long-tailed chinchilla (Chinchilla lanigera) under natural photoperiod.

Chinchilla spp. is a South American hystricomorph rodent genus currently considered almost extinct in the wild. The high quality of chinchilla fur motivated the harvesting of chinchillas for the fur market. Reproductive biology advances come from studies on commercially exploited animals, especially Chinchilla lanigera. We studied seasonal variation of urinary androgen metabolites, sperm concentration and sperm functional activity in males of domestic Chinchilla lanigera under natural photoperiod. In Córdoba city (31° S-64° W; Argentina), within the same latitudes as those of the historic Andean distribution (tropical deserts; 15°-34° S), domestic males (n=7) were studied in May (autumn), August (winter), November (spring), and February (summer). Urine was seasonally collected (over 24h; once for season, 4 in total) to measure urinary androgen metabolites (RIA), before semen collection by electroejaculation. The results indicated that although testicular volume (relative to body weight) and values of sperm functional activity did not show seasonal changes, a seasonal variation in androgen excretion was detected, with the highest values occurring during "short" light/dark cycles (autumn-winter). In addition, viable spermatozoa with intact acrosome mean values during winter-spring were higher than in autumn or summer. This study provides information that might contribute to the assessment of testicular activity in male chinchilla subjected to genetic selection in the fur industry. In addition, since domestic chinchilla still share some genomic characteristics with their counterparts in the wild, results presented may alsocontribute to ex situ breeding program of endangered chinchilla. In conclusion, natural photoperiod cycle affects testicular activity in domestic chinchilla.

Sexual hormone fluctuation in chinchillas.

The data about chinchilla (Chinchilla laniger) reproduction are limited and in some cases discordant. The aim of this study was to monitor the sexual hormone fluctuation by fecal progesterone level and colpocytology analysis by vaginal smears in order to evaluate the different phases of the oestrus cycle. Twenty-four non pregnant chinchillas aged from 1 to 4 years old and subdivided in three groups were monitored. In contrast with findings reported in other study, the high values of progesterone recorded in autumn suggested the presence of a ciclicity also in this period. The data indicate that chinchilla presents a continuous cycle.

Adrenal activity and anxiety-like behavior in fur-chewing chinchillas (Chinchilla lanigera).

Due to its complexity, in combination with a lack of scientific reports, fur-chewing became one of the most challenging behavioral problems common to captive chinchillas. In the last years, the hypothesis that fur-chewing is an abnormal repetitive behavior and that stress plays a role in its development and performance has arisen. Here, we investigated whether a relationship existed between the expression and intensity of fur-chewing behavior, elevated urinary cortisol excretion and anxiety-related behaviors. Specifically, we evaluated the following parameters in behaviorally normal and fur-chewing animals of both sexes: (1) mean concentrations of urinary cortisol metabolites and (2) anxiety-like behavior in an elevated plus-maze test. Urinary cortisol metabolites were higher only in females that expressed the most severe form of the fur-chewing behavior (P≤0.05). Likewise, only fur-chewing females exhibited increased (P≤0.05) anxiety-like behaviors associated with the elevated plus-maze test. Overall, these data provided additional evidence to support the concept that fur-chewing is a manifestation of physiological stress in chinchilla, and that a female sex bias exists in the development of this abnormal behavior.

Surtos de intoxicação por salinomicina em chinchilas (Chinchilla lanigera) / Outbreaks of salinomycin toxicosis in Chinchillas (Chinchilla lanigera)

Quatro surtos de intoxicação por salinomicina são descrito em chinchilas de três municípios do Estado do Rio Grande do Sul. Uma semana após a ingestão de ração contendo 37 ppm de salinomicina, aproximadamente duas mil chinchilas de quatro fazendas expostas diminuíram o consumo da ração. Quatrocentos e vinte sete chinchilas demonstraram apatia. Dessas, duzentos e setenta e sete desenvolveram decúbito esternal e lateral, dispneia e coma, seguidos de morte. As primeiras mortes ocorreram oito dias após a ingestão da ração. A evolução dos sinais clínicos até a morte ou eutanásia foi de 2-5 dias. Os exames bioquímicos do soro sanguíneo em quatro chinchilas revelaram níveis aumentados da alanina aminotransferase, aspartato transaminase, fosfatase alcalina, creatina cinase, glicose, triglicerídeos e colesterol total. Quarenta e cinco chinchilas foram submetidas à necropsia. Os achados macroscópicos consistiam de marcada lipidose hepática em todas as chinchilas necropsiadas; fetos em estado de decomposição em doze chinchilas que estavam prenhes. Microscopicamente, múltiplas fibras musculares esqueléticas estavam hipereosinofílicas, tumefeitas e com perda das estriações. Nas chinchilas que sobreviveram por mais dias era possível observar segmentos fragmentados de miofibras afetadas (necrose flocular) e regeneração de miofibras. No fígado foi observada marcada degeneração gordurosa. Não foram observadas anormalidades microscópicas nos demais órgãos analisados. Análises à procura de aflatoxinas, resíduos de pesticidas e isolamento bacteriano foram negativos. A análise da ração por cromatografia líquida revelou 37ppm de salinomicina na ração. A ração suspeita foi administrada a 12 chinchilas, três das quais (25 por cento) morreram apresentando lesões semelhantes às observadas nas chinchilas com a doença natural. O diagnóstico de intoxicação por salinomicina foi baseado na epidemiologia, lesões histológicas características e na presença de salinomicina na ração administrada nas quatro criações envolvidas.

Four outbreaks of ionophore toxicosis are described in chinchillas from four commercial farms located in three municipalities in the state of Rio Grande do Sul, southern Brazil. Approximately 2,000 chinchillas showed decrease in food intake one week after start ingesting a ration containing 37 ppm of salinomycin. Four hundred and twenty seven chinchillas showed apathy. Of those 277 develop sternal and lateral recumbence, dyspnea and coma followed by death. First deaths occurred eight days after the start on the salinomycin containing ration; clinical course was 2-5 days. Serum chemistry carried out in four chinchillas revealed increased levels of alanine transaminase, aspartate transaminase, alkaline phosphatase, creatinenin kinase, glucose, triglicerids and total cholesterol. Forty five affected chinchillas were necropsied; consistent necropsy findings were marked hepatic lipidosis; additionally twelve pregnant chinchillas had dead decomposing fetuses. Microscopically skeletal muscles had multifocally swollen hypereosinophilic myofibers with loss of cross striations. In those chinchillas that survived longer than a few days, microscopic features in the skeletal muscle included segmental fragmentation of dead fibers (floccular necrosis) and myofiber regeneration. Marked fatty degeneration was observed in the livers of all affected chinchillas. No microscopic changes were observed in other organs. Chemical analysis in the feed consumed by the chinchillas did not detect aflatoxins or pesticides residues; bacterial culture performed in samples of liver and intestinal contents from necropsied chinchillas yielded no significant bacterial growth. Analysis by thin layer chromatography performed in the ration consumed by the chinchillas detected 37 ppm of salinomycin. The suspected ration was fed to 12 chinchillas three of which (25 percent) died with similar lesions to those observed in the natural cases. The diagnosis of salinomycin toxicosis was based in the epidemiology, histology of the lesions, on the detection of significant amounts of salinomycin in the ration used to feed the chinchillas in the four involved farms and on the reproduction of disease by feeding the suspected ration to susceptible chinchillas.

The pituitary of non-pregnant and pregnant viscachas (Lagostomus maximus maximus): a comparative study by immunohistochemistry and morphometric analysis.

The neuroendocrine hypothalamic-pituitary axis undergoes morphological and biochemical changes throughout gestation. In viscacha, pregnancy lasts approximately 154 days, and three stages can be described: early, mid- and late pregnancy. The aim of this work was to study the pituitary LH-gonadotrophs, FSH-gonadotrophs, somatotrophs, corticotrophs and thyrotrophs of non-pregnant and pregnant adult viscachas by immunohistochemistry and morphometric analysis. Immunopositive percentage area (%IA), cell percentage in the pars distalis (%PDC), number of cells per reference area (n°cell/RA), and major cellular (MCD) and nuclear (ND) diameters were analyzed. The different cell populations showed a well-defined morphology, immunolabeling patterns and regionalization in the pars distalis (PD). In the early pregnancy of animals the morphometric analysis demonstrated that %IA, %PDC and n°cell/RA increased in the FSH-gonadotrophs and decreased in the somatotrophs in relation to non-pregnant animals. In mid-pregnancy, there was an increase in %IA, %PDC, and n°cell/RA of LH-gonadotrophs, FSH-gonadotrophs, somatotrophs, and thyrotrophs. The MCD of LH-gonadotrophs and FSH-gonadotrophs increased. In late pregnancy, the %IA, %PDC and n°cell/RA of LH-gonadotrophs, FSH-gonadotrophs, somatotrophs and corticotrophs decreased whereas the values of the thyrotrophs remained constant. The MCD of LH-gonadotrophs, FSH-gonadotrophs and corticotrophs decreased. No significant changes were observed in the ND of the studied cell types. In conclusion, this work provides evidence for histological and morphometric changes in the different cell types of the pituitary PD in viscachas during pregnancy, probably according to the requirements of this physiological stage.

Effects of cochlear ablation on amino acid concentrations in the chinchilla posteroventral cochlear nucleus, as compared to rat.

Using a microchemical approach, we measured changes of amino acid concentrations in the chinchilla caudal posteroventral cochlear nucleus (PVCN) after cochlear ablation to determine to what extent slow decreases of glutamate and aspartate concentrations after carboplatin treatment resulted from slower effects of cochlear damage in chinchillas than in rats and guinea pigs, as opposed to effects of carboplatin treatment being slower than those of cochlear ablation. Our results indicate that both factors are involved: decreases of glutamate and aspartate concentrations after cochlear ablation are much slower in chinchillas than in rats and guinea pigs, but they are much faster than the decreases after carboplatin treatment. Further, aspartate and glutamate concentrations in the chinchilla caudal PVCN decreased by larger amounts after cochlear ablation than in rats or guinea pigs, and there was a transient increase of aspartate concentration at short survival times. Detailed mapping of amino acid concentrations in the PVCN of a chinchilla with 1 month survival after cochlear ablation and a rat with 7 days' survival indicated that the reductions of glutamate and aspartate occurred throughout the PVCN but were somewhat larger in ventral and caudal parts in chinchilla. Any decreases in the adjacent granular region were very small. There were also sustained bilateral decreases in concentrations of other amino acids, notably GABA and glycine, in the caudal PVCN of cochlea-ablated chinchillas but not rats. The effects of cochlear ablation on the concentrations of most of these other amino acids in chinchilla caudal PVCN differed from those of carboplatin treatment. Thus, although a major effect of auditory nerve damage on the cochlear nucleus-decreases of glutamate and aspartate concentrations-occurs across species and types of lesions, the details of timing and magnitude and the effects on other amino acids can vary greatly.

Delayed mitochondrial dysfunction in apoptotic hair cells in chinchilla cochleae following exposure to impulse noise.

Apoptotic death of hair cells (HCs) in the cochlea has been found following exposure to intense noise. The current study was designed to examine the mitochondrial energetic function of HCs during the course of noise-induced apoptosis. Two aspects of the mitochondrial energetic function, succinate dehydrogenase (SDH) activity and mitochondrial membrane potential (MMP), were examined in HCs of chinchilla cochleae following exposure to a series of 75 pairs of impulse noises at 155 dB pSPL. The results showed that nuclear condensation and uptake of propidium iodide or trypan blue appeared at 10 min after the noise exposure, indicating a rapid progression of HC apoptosis. However, SDH activity was preserved at this time point. As the time elapsed (1 hr or 24 hrs) after the noise exposure, all newly-generated apoptotic HCs showed strong SDH activity, indicating the preservation of SDH activity during the course of apoptosis. Examination of MMP with rhodamine 123 staining revealed that MMP was sustained in the apoptotic HCs having mild nuclear condensation, even after the occurrence of cell membrane leakage. MMP was reduced with further progression of nuclear condensation. These results suggest the presence of a delayed mitochondrial dysfunction in apoptotic HCs following exposure to intense noise.

Noninvasive monitoring of ovarian endocrine activity in the chinchilla (Chinchilla lanigera).

Reproductive endocrinology information is limited for Chinchilla lanigera, a South American species characterized by extremely long gestation and estrus cycle compared with others rodents. This study was designed to validate a non-invasive technique for monitoring ovarian endocrine activity. Animals were exposed indoors to natural photoperiod (31 degrees S-64 degrees W, Argentina); temperature range: 17-26 degrees C, with food and water ad libitum. Radiolabelled infusion (n=4): (3)H-estradiol ((3)H-E(2)) and (14)C-progesterone ((14)C-P(4)) were injected (i.p). Biochemical validation: HPLC-UV detector was employed to determine natural steroids in urine and fecal extracts and to determine immunoreactive metabolites. Physiological validation: (1) pregnancy (n=5): body weight and urinary and fecal steroidal metabolites were measured until birth; (2) Seasonality (n=9): urine and feces were collected in May, August, November, and February. Total (3)H-E(2) and (14)C-P(4) radioactivity recovered was 60.5+/-15.5 and 74.5+/-19.4%, respectively. After (3)H-E(2) injection, urinary radioactivity peaked at 7.0+/-0.6 hr; in contrast, urinary (14)C-P(4) excretion peaked at 44.0+/-4.0 hr (p=0.000). Peak radioactivity in feces occurred between 24-48 hr for both hormones. Several correlations were detected during pregnancy between body weight vs. urinary progestagens/day (r=0.44, p<0.03); vs. urinary progestagens/creatinine (r=0.73, p=2.9 x 10(-5)); vs. urinary estrogens/day (r=0.74; p<0.2); and vs. urinary estrogens/creatinine (r=0.74; p<2.0 x 10(-5)). On the other hand, urinary and fecal progestagen excretion exhibited significant seasonal fluctuations and urinary estrogen concentrations showed a similar pattern (p=0.062 for winter-spring vs. summer-autumn). This methodology proved to be useful for monitoring ovary endocrine activity in urine of chinchilla female.

A member of the cathelicidin family of antimicrobial peptides is produced in the upper airway of the chinchilla and its mRNA expression is altered by common viral and bacterial co-pathogens of otitis media.

Cationic antimicrobial peptides (AMPs), a component of the innate immune system, play a major role in defense of mucosal surfaces against a wide spectrum of microorganisms such as viral and bacterial co-pathogens of the polymicrobial disease otitis media (OM). To further understand the role of AMPs in OM, we cloned a cDNA encoding a cathelicidin homolog (cCRAMP) from upper respiratory tract (URT) mucosae of the chinchilla, the predominant host used to model experimental OM. Recombinant cCRAMP exhibited alpha-helical secondary structure and killed the three main bacterial pathogens of OM. In situ hybridization showed cCRAMP mRNA production in epithelium of the chinchilla Eustachian tube and RT-PCR was used to amplify cCRAMP mRNA from several other tissues of the chinchilla URT. Quantitative RT-PCR analysis of chinchilla middle ear epithelial cells (CMEEs) incubated with either viral (influenza A virus, adenovirus, or RSV) or bacterial (nontypeable H. influenzae, M. catarrhalis, or S. pneumoniae) pathogens associated with OM demonstrated distinct microbe-specific patterns of altered expression. Collectively, these data showed that viruses and bacteria modulate AMP messages in the URT, which likely contributes to the disease course of OM.

The chinchilla microdialysis model for the study of antibiotic distribution to middle ear fluid.

In cases of slow or limited penetration of an antibiotic to the site of infection such as in acute otitis media (the middle ear), plasma levels of the agent may not reflect the concentrations that are relevant in determining clinical outcome. There is a need for a model that allows prediction of the time-course of unbound, pharmacologically active drug levels in middle ear fluid (MEF). This article introduces microdialysis as a sampling tool to measure unbound antibiotic concentrations in the MEF of the chinchilla, and briefly summarizes the results of studies of MEF penetration of a cephalosporin, a macrolide, and a ketolide antibiotic using this technique. The general concurrence of preliminary results of the chinchilla studies with clinical findings suggests that the chinchilla microdialysis model may be useful in predicting efficacy in patients.

Assessment of urine and fecal testosterone metabolite excretion in Chinchilla lanigera males.

Endemic chinchilla (Chinchilla spp.) populations are nearly extinct in the wild (South America). In captive animals (Chinchilla lanigera and C. brevicaudata), reproduction is characterized by poor fertility and limited by seasonal breeding patterns. Techniques applied for studying male reproductive physiology in these species are often invasive and stressful (i.e. repeated blood sampling for sexual steroids analysis). To evaluate endocrine testicular function, the present experiments were designed to (a) determine the main route of testosterone excretion (14C-testosterone infusion in four males); (b) validate urine and fecal testosterone metabolite measurements (HPLC was used to separate metabolites and immunoreactivity was assessed in all metabolites using a commercial testosterone radioimmunoassay, and parallelism, accuracy and precision tests were conducted to validate the immunoassay); and (c) investigate the biological relevance of the techniques applied (quantification of testosterone metabolite excretion into urine and feces from five males injected with hCG and comparison between 10 males and 10 females). Radiolabelled metabolites of 14C-testosterone were excreted, 84.7+/-4.2 % in urine and 15.2+/-3.9 % in feces. A total of 82.7+/-4.2% of urinary and 45.7+/-13.6% of fecal radioactivity was excreted over the first 24 h period post-infusion (metabolite concentration peaked at 8.2+/-2.5 h and 22.0+/-7.0 h, respectively). Several urinary and fecal androgen metabolites were separated by HPLC but only fecal metabolites were associated with native testosterone; however, there was immunoreactivity in more than one metabolite derived from 14C-testosterone. After hCG administration, an increase in androgen metabolite excretion was observed (p<0.05). Males excreted greater amounts daily of urinary androgen metabolites as compared with females (p<0.05); this difference was not evident in feces. Results of the present study indicate that the procedure used is a reliable and non-invasive method to repeatedly monitor variations in testicular endocrine activity in this species. It can be a useful tool that would help ensure the survival of the wild populations as well as to provide the basis for a more efficient use by the fur industry.

A non-invasive method for assessing adrenal activity in the chinchilla (Chinchilla lanigera).

The Chinchilla is a rodent that was once abundant in the central Andes of South America. Excessive hunting for fur greatly reduced its distribution at the beginning of the twentieth century, and today Chinchilla species are nearly extinct in the wild. Although protected, wild populations of chinchilla are still declining. In general, this species has received little research attention and its biology is poorly understood. Improvements in captive breeding, husbandry, and genetic management are needed to ensure the conservation of the species. In this study, a noninvasive corticosteroid hormone monitoring technique was validated for use in Chinchilla lanigera. Two male domestic chinchillas were administered 3H-corticosterone (i.m.) to determine the time course and relative proportion of urinary and fecal steroid metabolites. Most radioactivity was detected in urine and feces 5-10 and approximately 30 h post-isotope administration, respectively. Corticosteroid immunoreactivity was assessed by corticosterone radioimmunoassay (RIA) and cortisol enzyme immunoassay (EIA). High-pressure liquid chromatography (HPLC) separation of corticosteroid metabolites in unprocessed urine revealed the presence of highly polar corticosteroid metabolites, but after enzymatic hydrolysis and diethyl ether extraction, most immunoreactivity co-eluted with unconjugated cortisol. A 'cause-and-effect' relationship between the administration of exogenous adrenocorticotrophic hormone (ACTH), and the appearance of increased urinary corticosteroid metabolites demonstrated the physiological relevance of these measures for evaluating adrenal status in male chinchillas. From a conservation perspective, these methods can aid in situ and ex situ initiatives designed to evaluate how environmental conditions and management strategies affect overall animal health, well-being and reproduction.

The nutrition of the chinchilla as a companion animal--basic data, influences and dependences.

This contribution is meant to obtain basic data for feeding chinchillas (ingestion behaviour, feed and water intake) kept as companion animals. The chinchillas ingested more than 70% of their total feed intake during the dark phase (highest level of activity between 9:00 pm and 7:00 am). Daily amounts of feed intake varied between 2.5 (fresh grass) or 2.6 (hay) and 5.5 (pelleted complete diet) g of dry matter per 100 g of body weight. An offered mixed feed based on native components led to a selection of individual ingredients (high palatability: carob, beet pulp, sunflower seeds). The chinchillas' daily water intake varied between 30 (mixed feed in briquette form) and 40 ml (alfalfa cubes) and amounted on average between 1.5 and 3 ml/g of dry matter. Compared with rabbits or guinea-pigs, the chinchillas generally showed noticeable differences (rhythm of feed intake, palatability of individual ingredients, capacity for digestion, etc.) which must be considered in order to optimize the nutrition of this species.

Tracking the tissue distribution of marker dye following intranasal delivery in mice and chinchillas: a multifactorial analysis of parameters affecting nasal retention.

The combined mucosal surface area is vast and represents the primary site of entry of most pathogens to the respiratory, gastrointestinal (GI), and urogenital tracts. It is recognized that vaccines delivered parenterally typically only induce weak mucosal immune responses and that by targeting the mucosal immune system protective antibodies and effector lymphocytes could be induced at the primary site of infection. In the present study, we have evaluated an extensive set of conditions required to ensure maximum retention of intranasally administered substances within the nasal cavity in the chinchilla and murine hosts. We report here that many parameters, such as anaesthesia, position of animal during and post delivery, and dosing schedule, must be optimized in concert with each other and that results from one species cannot be extrapolated directly to another animal model.

Expression of heregulin and ErbB/Her receptors in adult chinchilla cochlear and vestibular sensory epithelium.

Immunolabeling of heregulin, a growth factor that enhances cell proliferation in damaged utricles, and one of its binding receptors, ErbB-2, has been briefly described in the P3 rat cochlea and utricle [Zheng et al. (1999) J. Neurocytol. 28, 901-912]. However, little is known about the distribution of heregulin and its three binding receptors in adult animals. Here we describe the immunolabeling patterns for heregulin, ErbB-2, ErbB-3 and ErB-4 in the cochlea, spiral ganglion, utricle and saccule of the adult chinchilla using confocal microscopy. Heregulin immunolabeling was intense along the apical pole of Deiters cells and Hensen cells and along the membrane of supporting cells of the utricle and saccule; light immunolabeling was present in the outer layer of the spiral prominence and cytoplasm of spiral ganglion neurons. In the cochlea, intense to moderate ErbB-2 immunolabeling was evident in the cytoplasm of pillar cells, outer hair cells (OHCs), border cells, stria vascularis and spiral ligament; moderate ErbB-2 immunolabeling was present in the cytoplasm of the hair cell and supporting cell layers of the utricle and saccule. In the cochlea, light ErbB-3 immunolabeling was present in the inner hair cells, OHCs, marginal and intermediate cell layers of the stria vascularis and spiral ganglion neurons; moderate ErbB-3 immunolabeling was present in the cytoplasm of hair cells and supporting cells of the utricle and saccule. In the cochlea, utricle and saccule, ErbB-4 immunolabeling was intense in the nuclei and light to moderate in the cytoplasm and membrane of sensory cells and supporting cells. These results suggest that heregulin acting through ErbB receptors and various receptor complexes may play an important role in cell proliferation and survival in the cochlea and vestibular system.