Ovarian nesfatin-1 in Hemidactylus flaviviridis: Reproductive phase-dependent expression, role and hormonal regulation.

Nesfatin-1 has recently emerged as a modulator of ovarian functions in mammals. Studies in non-mammalian vertebrates, though limited and majorly restricted to fishes, have evidenced a role of this peptide in the regulation of ovarian steroidogenesis and oocyte maturation. Interestingly, nesfatin-1 remains completely unexplored in reptiles. Therefore, the present study aimed to identify nesfatin-1 and elucidate its role and regulation in the ovary of Hemidactylus flaviviridis. Ovarian expression of nucb2/nesfatin-1 was highest during late recrudescence and breeding while it was lowest during regression. Follicular stage-dependent expression analysis showed significantly high expression of nucb2/nesfatin-1 in previtellogenic follicles. Further, in vitro treatment of recrudescent wall lizard ovaries with nesfatin-1 resulted in increased expression of anti-apoptotic gene, bcl-2, along with a concomitant decline in the pro-apoptotic gene, caspase-3. In addition, proliferation/differentiation markers like scf, c-kit, pcna, and bmp-15 were stimulated in ovaries incubated with the peptide. Ovarian steroidogenesis was also positively influenced by nesfatin-1 as treatment with the peptide resulted in heightened star expression as well as increased estradiol and progesterone production. Also, all concentrations of nesfatin-1 stimulated glucose uptake and metabolism in wall lizard ovary. Our observations provide the first evidence of ovarian functions of nesfatin-1 in a reptile. Further, ovarian nucb2/nesfatin-1 was differentially regulated by gonadotropin and sex steroids wherein its expression was stimulated by dihydrotestosterone (DHT) and 17ß-estradiol (E2) but inhibited by follicle-stimulating hormone (FSH). In summary, this is the first report of the presence, reproductive stage-dependent expression, role, and regulation of ovarian nucb2/nesfatin-1 in H. flaviviridis.

Nesfatin-1 in a reptile: its role and hormonal regulation in wall lizard testis.

Nesfatin-1 is a pleiotropic hormone implicated in various physiological functions including reproduction. Studies though limited, have established an important role of the peptide in regulation of testicular functions in mammals and fishes. However, role of nesfatin-1 in regulation of spermatogenesis and testicular steroidogenesis remains completely unexplored in reptiles. Therefore, present study aimed to develop an insight into reproductive phase-dependent testicular expression, function and regulation of nucb2/nesfatin-1 in a reptile, Hemidactylus flaviviridis. Expression of nucb2/nesfatin-1 in testis of wall lizard varied significantly depending upon reproductive phase, being highest in the active phase while lowest during regressed phase. Further, in vitro treatment of wall lizard testis with nesfatin-1 showed a concentration- and time-dependent stimulatory effect of the peptide on expression of cell proliferation and differentiation markers like scf, c-kit and pcna suggesting a spermatogenic role of nesfatin-1 in wall lizard. Also, nesfatin-1 stimulated the anti-apoptotic marker, bcl-2 while inhibited the apoptotic marker, caspase-3, suggesting its role as an inhibitor of apoptosis of testicular cells. Further, treatment with nesfatin-1 resulted in significantly higher expression of star along with a concomitant increase in testosterone production by the lizard testis. The present study also demonstrates hormonal regulation of testicular nucb2/nesfatin-1 wherein follicle-stimulating hormone (FSH) inhibited while sex steroids like dihydrotestosterone (DHT) and 17ß-estradiol-3-benzoate (E2) stimulated the mRNA expression of nesfatin-1. Observations from the current study for the first time provide comprehensive evidence of spermatogenic and steroidogenic role of nesfatin-1 as well as its hormonal regulation in the testis of a reptile, H. flaviviridis.

Reproductive phase-dependent and sexually dimorphic expression of leptin and its receptor in different parts of brain of spotted snakehead Channa punctata.

The reproductive phase-wise leptin (lep) and its receptor (lepr) expression in different parts of the brain of adult male and female spotted snakehead Channa punctata reveals sexual dimorphism in the brain leptin system. In anterior, middle and posterior parts of the brain of males, a maximum lep was observed in resting, spawning and postspawning reproductive phases, respectively. In females, a high level of lep was seen during the preparatory phase in the anterior brain, preparatory and postspawning phases in the middle brain and resting and postspawning phases in the posterior brain. Nonetheless, the transcript level of lepr was recorded highest during the spawning phase, irrespective of sex and region of the brain. Regardless of the reproductive state of fishes, lep and lepr were seen considerably high in middle and posterior parts of male brain than that of female, implying the involvement of factors other than sex steroids for sex-related variation in the leptin system in these regions of the brain. Nonetheless, no sex difference was evidenced in the expression of either ligand or its receptor in the anterior brain. In summary, the presence of lep and lepr in different regions of the brain and variation in their expression depending on sex and reproductive phases raise the possibility of pivotal actions of leptin in influencing neuronal circuitry and thereby reproductive functions.

Crosstalk between reproductive and immune systems: the teleostean perspective.

The bidirectional interaction between the hypothalamic-pituitary-gonadal (HPG) axis and the immune system plays a crucial role in the adaptation of an organism to its environment, its survival and the continuance of a species. Nonetheless, very little is known about this interaction among teleost, the largest group of extant vertebrates. Fishes being seasonal breeders, their immune system is exposed to seasonally changing levels of HPG hormones. On the contrary, the presence and infiltration of leukocytes, the expression of pattern recognition receptors as well as cytokines in gonads suggest their key role in teleostean gametogenesis as in the case of mammals. Moreover, the modulation of gametogenesis and steroidogenesis by lipopolysaccharide implicates the pathological significance of inflammation on reproduction. Thus, it is important to engage in the understanding of the interaction between these two important physiological systems, not only from a phylogenetic perspective but also due to the importance of fish as an important economic resource. In view of this, the authors have reviewed the crosstalk between the reproductive and immune systems in teleosts and tried to explore the importance of this interaction in their survival and reproductive fitness.

Exploring the effect of dihydrotestosterone on nucleotide-binding and oligomerization domain-like receptor expression in spotted snakehead Channa punctata (Bloch 1793).

Sex steroids are known to modulate immune responses and as a result many of the immune parameters in seasonally breeding organisms show reproductive-phase dependent variation. Androgens, the male sex steroids, are largely reported to be immunosuppressive. Together with other pattern recognition receptors, the nucleotide-binding and oligomerization domain-like receptors (NLRs) serve as intracellular sentinels and are essential to defense mechanisms. Interestingly, to date the transcriptional modulation of NLRs by androgens has not been explored. In the present study, we investigated the reproductive-phase dependent expression of NLRs in the male spotted snakehead Channa punctata. Furthermore, the effect of dihydrotestosterone (DHT) on NLR expression was studied. The expression of NLRs was observed to be most pronounced during the spawning phase of the fish, which is marked by the highest testosterone level. In vivo as well as in vitro studies showed the diverse effect of DHT on NLR expression depending on the duration and mode of treatment, as well as the immune tissue studied.

The role of non-ionizing electromagnetic radiation on female fertility: A review.

With increasing technological developments, exposure to non-ionizing radiations has become unavoidable as people cannot escape from electromagnetic field sources, such as Wi-Fi, electric wires, microwave oven, radio, telecommunication, bluetooth devices, etc. These radiations can be associated with increased health problems of the users. This review aims to determine the effects of non-ionizing electromagnetic radiations on female fertility. To date, several in vitro and in vivo studies unveiled that exposure to non-ionizing radiations brings about harmful effects on oocytes, ovarian follicles, endometrial tissue, estrous cycle, reproductive endocrine hormones, developing embryo, and fetal development in animal models. Non-ionizing radiation also upsurges the free radical load in the uterus and ovary, which leads to inhibition of cell growth and DNA disruptions. In conclusion, non-ionizing electromagnetic radiations can cause alterations in both germ cells as well as in their nourishing environment and also affect other female reproductive parameters that might lead to infertility.

Seasonality, sex-specificity and transcriptional regulation of hepatic leptin system in spotted snakehead Channa punctata.

The present study deals with sex-specific reproductive phase-dependent variation and sex steroids-induced transcriptional regulation of hepatic lep and lepr in nutritionally valuable spotted snakehead, Channa punctata. The data on seasonality reveals sex-specific variation in pattern of lep transcription where a high level was recorded during resting and postspawning quiescent phases in female while during resting and spawning phases in male. Unlike lep, lepr exhibited similar expression pattern along the reproductive phases in both the sexes. As compared to female, a three-fold higher expression of lep was detected in male during reproductively active phase only. However, no sexual dimorphism was evidenced in lepr either during active or quiescent phase. To explore the implication of sex steroids in regulation of leptin system, we correlated levels of plasma testosterone (T) and 17ß-estradiol (E2) with leptin system in males as well as females. Further, criss-cross in vivo and in vitro experiments with dihydrotestosterone (DHT) and E2 were conducted in male and female spotted snakehead. The leptin system was downregulated after DHT administration in both the sexes. However, with E2, a marked decrease was evidenced in male only. The sex-wise variable response of leptin system to sex steroids was validated by in vitro experiments wherein liver fragments from male and female fish were incubated individually with both the sex steroids. In conclusion, sex steroids modulate hepatic leptin system differentially depending on sex and reproductive state of spotted snakehead.

Testicular 25-hydroxycholesterol: An alternate substrate for steroidogenesis in reptiles.

The current study in wall lizards Hemidactylus flaviviridis was designed to ascertain that Leydig cells utilize testicular macrophage-derived 25-hydroxycholesterol (25-HC) for steroidogenesis. Leydig cells (LC) collected from regressed testes when incubated with 25-HC that was obtained from HPLC-eluted fraction of testicular macrophage-conditioned medium (TMCM), lyophilized and reconstituted in culture medium (0.5 µg/ml/well), produced considerably higher amount of testosterone. A similar observation was made when Leydig cells were incubated with varying concentrations of commercial 25-HC. Testosterone production by LC increased in a concentration-dependent manner. Taken together, it is evident that LC utilize 25-HC as a substrate for testosterone biosynthesis. To examine the gonadotropic regulation of steroid biosynthesis utilizing 25-HC as substrate, ovine follicle-stimulating hormone (FSH) that regulates both the testicular functions in lizards was used. Leydig cells were incubated with combinations of FSH and 25-HC as follows: 0 h FSH + 12 h 25-HC, 0 h 25-HC + 12 h FSH. As compared to respective controls, a marked increase in testosterone production was observed in response to FSH indicating that gonadotropin up-regulates uptake of 25-HC as a substrate for testosterone biosynthesis.

A comparative account of nesfatin-1 in vertebrates.

Nesfatin-1 was discovered as an anorexigenic peptide derived from proteolytic cleavage of the prepropeptide, nucleobindin 2 (NUCB2). It is widely expressed in central as well as peripheral tissues and is known to have pleiotropic effects such as regulation of feeding, reproduction, cardiovascular functions and maintenance of glucose homeostasis. In order to execute its multifaceted role, nesfatin-1 employs diverse signaling pathways though its receptor has not been identified till date. Further, nesfatin-1 is reported to be under the regulatory effect of feeding state, nutritional status as well as several metabolic and reproductive hormones. This peptide has also been associated with variety of human diseases, especially metabolic, reproductive, cardiovascular and mental disorders. The current review is aimed to present a consolidated picture and highlight lacunae for further investigation in order to develop a deeper comprehensive understanding on physiological significance of nesfatin-1 in vertebrates.

In silico analysis, seasonal variation and gonadotropic regulation of jag1 and its receptor notch1 in testis of spotted snakehead Channa punctatus.

The present study in seasonally breeding spotted snakehead Channa punctatus, for the first time in nonmammalian vertebrates, demonstrated correlation between reproductive phase-dependent testicular expression of ligand Jag1/receptor Notch1 and spermatogenic events. Testicular transcriptome sequencing data from our earlier study in C. punctatus was used in the present study to select the best transcript for jag1 (cpjag1) and notch1 (cpnotch1). The transcripts cpjag1 and cpnotch1 encoded full-length putative proteins of 1215 (cpJag1) and 2475 (cpNotch1) amino acids, respectively. A marked homology in the extracellular domains of Jag1 and Notch1 was observed following their alignment with respective proteins from different vertebrates, suggesting conservation in ligand-receptor interaction in C. punctatus. Both cpJag1 and cpNotch1 showed phylogenetic closeness with their teleostean counterparts, especially with that of Perciformes. Temporal expression of cpjag1 and cpnotch1 in testis depending on reproductive phases showed an appreciably high expression during spermatogenically inactive resting and postspawning phases when seminiferous lobules consisted of spermatogonial stem cells and undifferentiated spermatogonia. Their expression sharply declined during spermatogenically active preparatory and spawning phases. It appears that involvement of cpjag1/cpnotch1 is restricted to inactive phases when spermatogonial stem cells renew themselves and replenish undifferentiated spermatogonia. This assumption is ascertained by an experimental study in which high level of testicular cpjag1/cpnotch1 expression in control fish of resting phase markedly decreased after administration of human chorionic gonadotropin that is known to induce proliferation and differentiation of spermatogonia and spawning of spermatozoa.

In silico analysis, temporal expression and gonadotropic regulation of receptors for follicle-stimulating hormone and luteinizing hormone in testis of spotted snakehead Channa punctata.

This study in spotted snakehead Channa punctata was aimed to develop a comprehensive understanding of testicular gonadotropin receptors, from their sequence characterization, temporal expression to gonadotropic regulation, in seasonally breeding teleosts. A single form of follicle-stimulating hormone receptor (cpfshra) and luteinizing hormone/choriogonadotropin receptor (cplhcgr), was identified from testicular transcriptome data of C. punctata. Although deduced full-length protein sequence for cpFshra (694 amino acids) and cpLhcgr (691 amino acids) showed homology with their counterparts of other vertebrates, multiple insertion-deletion-substitution of residues suggest marked alterations in their structure and ligand specificity. The absolute quantification of testicular cpfshra and cplhcgr was estimated along the reproductive cycle following real-time PCR. The temporal expression profile showed highest testicular expression of both the gonadotropin receptors during resting phase. Their expression progressively decreased during preparatory and spawning phases concomitant with spermatogonial proliferation and differentiation and spermiogenesis. However, levels of cpfshra and cplhcgr sharply increased during post-spawning when seminiferous lobules were largely devoid of germ cells. To explore gonadotropic regulation of testicular cpfshra and cplhcgr, one group of fish of resting phase was administered with single dose of human chorionic gonadotropin (hCG; 5,000 IU/kg body mass) on day 0 and sacrificed on day 3 and day 5, while another group receiving two injections of hCG (day 0 and day 7) was sacrificed on day 14. The expression pattern of testicular gonadotropin receptors in hCG-treated fish sacrificed after 3, 5 and 14 days was similar to that of preparatory, spawning and postspawning phases, respectively. Likewise, testicular histology of hCG-treated fish sacrificed on day 3, day 5 and day 14 was comparable with that of preparatory, early spawning and late spawning phases, respectively. In light of the fact that gonadotropin receptors are largely expressed on somatic cells, an apparent decrease in testicular cpfshra and cplhcgr levels during preparatory and spawning phases or after 3 and 5 days from first hCG injection might not be due to downregulation of their expression. Rather, this could be due to dilution of somatic cell mRNA by large amount of germ cell mRNA. To verify this assumption, effect of hCG on plasma level of androgens was investigated employing enzyme-linked immunosorbent assay. A marked increase in plasma level of testosterone and 11-ketotestosterone was observed after hCG treatment in C. punctata. This would have been possible only when hCG upregulated the expression of testicular gonadotropin receptors.

Repertoire of bone morphogenetic proteins and growth/differentiation factors in ovary of the Indian wall lizard (Hemidactylus flaviviridis) with emphasis on differential expression and gonadotropic regulation of bmp15 and gdf9.

Analysis of ovarian transcriptome of Indian wall lizard demonstrates the existence of several bone morphogenetic proteins (bmp1, 2, 3, 3b, 7, 8, 15) and growth/differentiation factors (gdf5, 9) for the first time in reptilian ovary. The characterization of putative full-length/partial protein sequences of BMPs (BMP2, 3, 3b, 7, 15) and GDF9 showed high homology of their TGF-ß domain with that of other vertebrates while BMP1 bore homology to zinc-dependent metalloprotease. Phylogenetic analyses showed clustering of BMPs and GDF9 from wall lizards with that of squamates lying in close proximity to chelonia, crocodilia and aves. This study also correlates the expression of ovarian bmp15 and gdf9 with folliculogenesis. Level of bmp15 dramatically increased with the onset of follicular growth in early recrudescence and attained peak during late recrudescence whereas gdf9 sharply decreased during recrudescence as compared to regression. Nonetheless, expression of these growth factors decreased appreciably with the formation of vitellogenic follicle during breeding phase. Ovarian expression of bmp15 and gdf9 appeared to be regulated by gonadotropin as bmp15 considerably increased while gdf9 decreased in parallel to follicular development after administration of 3 injections of FSH. Expression of both the growth factors declined with the prolongation of treatment that led to formation of early/late vitellogenic follicle. Our in vitro study revealed stimulatory effect of FSH on expression of bmp15 and gdf9 in early growing, previtellogenic and early vitellogenic follicles. In light of in vitro results, FSH-induced in vivo decline in gene expression seems to be due to some other FSH-induced factor.

Purification and identification of 25-hydroxycholesterol in a reptile: Seasonal variation and hormonal regulation.

The present in vitro study, for the first time, demonstrates the production of 25-hydroxycholestrol (25-HC) by testicular macrophages of a non-mammalian vertebrate. The ether extracts of testicular macrophage-conditioned medium (TMCM) were fractionated on a C18 reversed phase high-performance liquid chromatography (HPLC) column using methanol as the mobile phase. The mass spectrometry (MS) fragmentation pattern of HPLC-purified 25-HC was found to be identical to that of authentic 25-HC. Further, a significant seasonal variation in 25-HC concentration was observed with maximal level in regressed and minimal during breeding phase. To understand the hormonal control of 25-HC production, testicular macrophages from regressed phase testes were incubated with 0.5µg/ml of ovine follicle stimulating hormone (FSH) and 0.1, 1 and 10µg/ml of testosterone (T). FSH considerably enhanced 25-HC production by testicular macrophages. In contrast, T markedly inhibited 25-HC production in a dose-dependent manner. In addition, T significantly inhibited FSH-induced 25-HC production, though pre-treatment with T was more effective as compared to post-treatment with T to FSH. Our findings on production, seasonal variation and hormonal control of 25-HC suggest the functional significance of 25-HC in the testis of reptiles.

Seasonality of reproduction in male spotted murrel Channa punctatus: correlation of environmental variables and plasma sex steroids with histological changes in testis.

The present study was undertaken to develop a comprehensive understanding of how environmental cues and sex steroids relate with cyclic changes in spermatogenesis in freshwater spotted snakehead Channa punctatus that is nutritious and economically important. The seasonal histological changes in testis and annual profile of gonadosomatic index (GSI) of C. punctatus delineated the testicular cycle into four phases: regressed (December-March), preparatory (April-June), spawning (July and August) and postspawning (September-November). Among environmental variables, correlation and regression analyses exhibited an important relationship between photoperiod and testicular weight while role of rainfall was seen confined to spawning. The seasonal profile of plasma sex steroids when correlated with cyclic changes in spermatogenesis in spotted snakehead, testosterone (T) seems to be involved in controlling the major events of spermatogenesis from renewal of stem cells to spawning of spermatozoa. Another important androgen prevalent in teleosts, 11-ketotestosterone (11-KT), was high during preparatory phase, suggesting that 11-KT in addition to T plays an important role in progression of spermatogenesis and spermiation in C. punctatus. However, 11-KT was not seen to be associated with milt production and release of spermatozoa during spawning. Plasma profile of estradiol-17ß (E2) during different reproductive phases revealed the involvement of E2 in repopulation of stem cells during postspawning phase and in maintaining quiescence of testis during regressed phase.

Radio frequency electromagnetic radiations interfere with the Leydig cell functions in-vitro.

A growing threat to male infertility has become a major concern for the human population due to the advent of modern technologies as a source of radiofrequency radiation (RFR). Since these technologies have become an integral part of our daily lives, thus, it becomes necessary to know the impression of such radiations on human health. In view of this, the current study aims to focus on the biological effects of radiofrequency electromagnetic radiations on mouse Leydig cell line (TM3) in a time-dependent manner. TM3 cells were exposed to RFR emitted from 4G cell phone and also exposed to a particular frequency of 1800 MHz and 2450 MHz from RFR exposure system. The cells were then evaluated for different parameters such as cell viability, cell proliferation, testosterone production, and ROS generation. A considerable reduction in the testosterone levels and proliferation rate of TM3 cells were observed at 120 min of exposure as compared to the control group in all exposure settings. Conversely, the intracellular ROS levels showed a significant rise at 60, 90 and 120 min of exposure in both mobile phone and 2450 MHz exposure groups. However, RFR treatment for different time durations (15, 30, 45, 60, 90, and 120 min) did not have significant effect on cell viability at any of the exposure condition (2450 MHz, 1800 MHz, and mobile phone radiation). Therefore, our findings concluded with the negative impact of radiofrequency electromagnetic radiations on Leydig cell's physiological functions, which could be a serious concern for male infertility. However, additional studies are required to determine the specific mechanism of RFR action as well as its long-term consequences.

Sexual dimorphism in NLR transcripts and its downstream signaling protein IL-1ꞵ in teleost Channa punctata (Bloch, 1793).

Nucleotide-binding oligomerization domain-like receptors (NOD-like receptors or NLRs) are a family of intracellular pattern recognition receptors (PRRs) that initiates as well as regulate inflammatory responses. NLRs are characterized by a centrally located nucleotide binding domain and a leucine rich repeat domain at the C-terminal responsible for the recognition of intracellular microbe-associated molecular patterns (MAMPs) and danger-associated molecular patterns (DAMPs). In the present study in adult spotted snakehead we have investigated the sex-dependent tissue distribution of NLRs known to be associated with inflammation in teleost namely NOD1, NOD2, NLRC3, NLRC5, and NLRX1. Further, the sexual dimorphism in the expression of NLR transcript as well as the pro-inflammatory protein IL-1ß was explored in fish under normal conditions, and in fish exposed to bacterial lipopolysaccharide (LPS). The NLRs show ubiquitous and constitutive expression in all the tissues. Moreover, a prominent disparity between males and females was observed in the basal expression of these genes in various tissues. The sexual dimorphism in NLR expression was also prominent when fish were exposed to LPS. Similarly, IL-1ß exhibited sexual dimorphism in both normal as well as LPS-exposed fish.

Genotoxic Risks to Male Reproductive Health from Radiofrequency Radiation.

During modern era, mobile phones, televisions, microwaves, radio, and wireless devices, etc., have become an integral part of our daily lifestyle. All these technologies employ radiofrequency (RF) waves and everyone is exposed to them, since they are widespread in the environment. The increasing risk of male infertility is a growing concern to the human population. Excessive and long-term exposure to non-ionizing radiation may cause genetic health effects on the male reproductive system which could be a primitive factor to induce cancer risk. With respect to the concerned aspect, many possible RFR induced genotoxic studies have been reported; however, reports are very contradictory and showed the possible effect on humans and animals. Thus, the present review is focusing on the genomic impact of the radiofrequency electromagnetic field (RF-EMF) underlying the male infertility issue. In this review, both in vitro and in vivo studies have been incorporated explaining the role of RFR on the male reproductive system. It includes RFR induced-DNA damage, micronuclei formation, chromosomal aberrations, SCE generation, etc. In addition, attention has also been paid to the ROS generation after radiofrequency radiation exposure showing a rise in oxidative stress, base adduct formation, sperm head DNA damage, or cross-linking problems between DNA & protein.

Molecular cloning, characterization and 3D modelling of spotted snakehead fbn1 C-terminal region encoding asprosin and expression analysis of fbn1.

The FBN1 gene encodes profibrillin protein that is cleaved by the enzyme furin to release fibrillin-1 and a glucogenic hormone, asprosin. Asprosin is implicated in diverse metabolic functions as well as pathological conditions in mammals. However, till date, there are no studies on asprosin in any non-mammalian vertebrate. In this study, we have retrieved the spotted snakehead Channa punctata fbn1 gene (ss fbn1) from the testicular transcriptome data and validated it. The transcript is predicted to encode 2817 amino acid long putative profibrillin protein. Amino acid sequence alignment of deduced ss profibrillin with human profibrillin revealed that the furin cleavage site in profibrillin is well conserved in C. punctata. Further, differential expression of ss fbn1 was observed in various tissues with the highest expression in gonads. Prominent expression of furin was also observed in the gonads suggesting the possibility of proteolytic cleavage of profibrillin protein and secretion of asprosin in C. punctata. In addition, the C-terminal of the fbn1 gene of C. punctata that codes for asprosin protein has been cloned. Using in silico approach, physicochemical properties of the putative ss asprosin were characterized and post-translational changes were predicted. The putative ss asprosin protein sequence is predicted to consist of 142 amino acid residues, with conserved glycosylation sites. Further, the 3D model of ss asprosin was predicted followed by MD (molecular dynamics) simulation for energy minimization. Thus, the current study, for the first time in non-mammalian vertebrates, predicts and characterizes the novel protein asprosin using in silico approach.

Significance of Complement Regulatory Protein Tetraspanins in the Male Reproductive System and Fertilization.

Fertilization is a very sophisticated and unique process involving several key steps resulting in a zygote's formation. Recent research has indicated that some immune system-related cell surface molecules (CD molecules from the tetraspanin superfamily) may have a role in fertilization. Extracellular vesicles are undeniably involved in a variety of cellular functions, including reproduction. Tetraspanin proteins identified in extracellular vesicles are now used mostly as markers; mounting evidence indicates that they also participate in cell targeting, cargo selection, and extracellular vesicle formation. Their significance and potential in mammalian reproduction are currently being studied extensively. Despite the fact that the current data did not establish any theory, the crucial function of tetraspanins in the fertilization process was not ruled out, and the specific role of tetraspanins is still unknown. In this review, we bring insight into the existing knowledge regarding the expression of tetraspanins in spermatozoa and seminal fluid and their role in gamete binding and fusion.