Goat Testis Research Articles

Characteristics of Sertoli cells in the ectopic and scrotal testes of unilateral cryptorchid West African dwarf goats.

Sertoli cell population and histology were studied in testes of unilateral cryptorchid goats. Histomorphological and electron microscopic techniques were employed. The results demonstrated that the number of Sertoli cell per testis was significantly (p < 0.05) higher in the descended scrotal testes of unilateral cryptorchid bucks compared with the right and left testes of the normal bucks, respectively. However there was no significant difference (p > 0.05) on comparing the number of Sertoli cells in the scrotal testes of the unilateral cryptorchid bucks with the paired testes of the normal bucks. Significant differences were not observed in the dimensions of the Sertoli cell nuclei in the retained, scrotal testes of the cryptorchids and the testes of the normal bucks. The Sertoli cells of the descended scrotal testes of the cryptorchid bucks and those of the normal bucks showed histological evidence of normal activity. Degeneration of Sertoli cells was observed in the retained testis of the cryptorchid bucks. The failure of one testis to descend most probably influenced compensatory hyperplasia of Sertoli cells and enhanced function of the cells in the descended testis of the unilateral cryptorchids.

113 Quantification of mRNA Expression for the Estrogen-Regulated Gene ISYNA1 in the Goat Testis

113 Quantification of mRNA Expression for the Estrogen-Regulated Gene ISYNA1 in the Goat Testis

230 EFFECT OF DIFFERENT CONCENTRATIONS OF GLIAL CELL LINE-DERIVED NEUROTROPHIC FACTOR ON EXPRESSION OF SELF-RENEWAL RELATED GENES IN GOAT (CAPRA HIRCUS) SPERMATOGONIAL STEM CELLS

Glial cell line-derived neurotrophic factor (GDNF), a member of the transforming growth factor-β superfamily produced by Sertoli cells, is essential for the self-renewal of spermatogonial stem cells in vivo. The present study evaluated the effects of different concentrations of GDNF (human recombinant expressed in Escherichia coli) on expression of some self-renewal related genes in spermatogonial stem cells (SSC). The SSC were isolated from prepubertal goat testes (3–6 months of age) by using double enzymatic digestion method and filtration through 80- and 60-µm nylon mesh filters. Further enrichment was achieved by differential plating on Datura stramonium agglutinin (DSA) lectin-coated dishes and Percoll density gradient centrifugation. The enriched cells were cultured on goat Sertoli cell feeder layer in DMEM + 10% fetal bovine serum at 37°C in a 5% CO2 incubator. Primary SSC colonies were formed within 7 to 10 days. These colonies were characterised through alkaline phosphatase and immunofluorescence staining on Day 10 for different SSC-specific protein markers. Colonies were found to be positive for DBA, THY1, PLZF, UCHL1, OCT-4, SOX2, and NANOG, and negative for c-Kit expression. These colonies were cultured for 15 days without or with supplementation of GDNF forming following groups: (1) without GDNF (control), (2) 10 ng mL–1 GDNF, (3) 20 ng mL–1 GDNF, and (4) 40 ng mL–1 GDNF. RNA was isolated from 100 colonies from 3 different trials on Day 15 of culture, and relative expression of different self-renewal related genes was determined by qRT-PCR. Relative mRNA abundance of PLZF was higher (P &lt; 0.05) following supplementation with 40 ng mL–1 GDNF than in other groups (i.e. control, 10 and 20 ng mL–1 GDNF). Expression of BCL6B and ID4 was found to be significantly higher (P &lt; 0.05) after supplementation of GDNF at all concentrations compared with the control group. Expression of UCHL1 was higher with addition of 20 and 40 ng mL–1 GDNF (P &lt; 0.05), whereas expression of THY1 was higher with supplementation of 10 ng mL–1 GDNF (P &lt; 0.05). In conclusion, GDNF was found to benefit expression of goat SSC candidate genes at a concentration of 40 ng mL–1.

Quantitative and cytological studies of interstitial (Leydig) cells in the scrotal and retained testes of unilateral cryptorchid West African Dwarf goats

Cryptorchidism is a common condition in West African Dwarf goats in Eastern Nigeria. The cytology and morphometry of interstitial cells were investigated using light and electron microscopy. Twenty adult male goats were used for the study. Ten of the goats were natural unilateral cryptorchids, while the remaining ten had fully descended testes and served as control. Significant hyperplasia of the interstitial cells were observed in scrotal and retained testes of the unilateral cryptorchid bucks compared to the control (normal) bucks. The number of interstitial cells per cross section of the testes was also significantly higher (P<0.05) in the contralateral scrotal testes of the unilateral cryptorchids (757.88±29.92) than in the testes of the normal bucks (677.30±25.92). In the contralateral scrotal testes of the unilateral cryptorchids, the interstitial cells had abundant cytoplasmic organelles, particularly mitochondria, Golgi apparatus and smooth endoplasmic reticulum, while the retained testes showed evidence of degeneration characterized by irregular nuclear profile, accumulation of nuclear heterochromatin, abundant lipid droplets, cytoplasmic vacuolations and focal intercellular dilations of interstitial cells. These results suggest that the intra-abdominal testes were significantly degenerated and the impairment was probably from exposure of the testes to higher abdominal temperature. However, the interstitial cells of the contralateral scrotal testes of the unilateral cryptorchid bucks showed normal histology and compensatory hyperplasia.

Investigation the mRNA expression of KISS1 and localization of kisspeptin in the testes of Shiba goats and its relationship with the puberty and steriodogenic enzymes

The objectives of the present study were to investigate the mRNA expression of KISS1 and localization of kisspeptin in the testes of Shiba goats. Additionally, its relationship with the puberty was investigated. Testes from prepubertal (1 month; n=3), and postpubertal Shiba goats (17±1.5 months; n=4) were collected by surgical castration. Plasma testosterone (T) and estradiol (E2) were measured just before the castration using radioimmunoassay. Testicular expression of mRNAs encoding KISS1, GPR54 and the following steroidogenic enzymes: cytochrome P450 side-chain cleavage (P450scc;CYP11A1 gene), 3beta-hydroxysteroid dehydrogenase (3βHSD; HSD3B1 gene), cytochrome P450, 17alpha-hydroxylase/17, 20 lyase (P450c17; CYP17 gene), and cytochrome P450 aromatase (P450arom; CYP19 gene) were quantified by real time PCR. The localization of kisspeptin in testes was determined by immunohistochemistry. Results revealed high plasma T and E2 in postpubertal goats compared to prepubertal goats. High significant increases (P<0.05) of mRNAs encoding KISS1, GPR54, CYP11A1, HSD3B1, CYP17, and CYP19 genes in the testes of postpubertal goats compared to prepubertal goats. In addition, kisspeptin was immunolocalized in Leydig cells of the testis. The staining was mild in the interstitial cells of the prepubertal goats while strong staining was found in the postpubertal goats. In conclusion: Leydig cells are responsible for kisspeptin production in the goat testis and this expression is puberty dependent.

Estrogen synthesis and secretion during postnatal testicular development in male goats: In situ aromatase mRNA expression

The role of testosterone in males has been extensively studied. However, studies in several species suggest that estrogens are also important for the development and function of male brain, bone, gonads as well as for spermatogenesis and sexual behavior. Data related to testicular P450-aromatase expression and estrogen receptors α and β in several species had been published. However, to our knowledge there is no information on testicular mRNA aromatase expression, estrogen synthesis, or estradiol-17β concentrations in the circulation of the male goat. Thus, the objective of this work was to conduct in vivo and in vitro studies related to estrogen synthesis and serum concentrations during the postnatal testicular development in goat bucks. To do this, circulating concentrations of estradiol-17β and testosterone were evaluated in male goat during the first year of life, starting at 15 days of age. Besides, in order to assess the estrogen-producing ability of testicles from prepubertal male goats (four months), testicular cells were incubated with tritiated androgen precursors (3H-androstenedione and 3H-testosterone) in the presence or absence of the P450-aromatase inhibitor formestane, using TLC to identify sex steroids. Finally, the expression and tissue distribution of P450-aromatase mRNA in the testis of 3–5 month old males was evaluated by in situ RT-PCR. Results indicate that estradiol-17β is present in the serum of male goats at 0.5 months of age, and that there are significant increases at 3 and 10 months of age. The in vitro synthesis of tritiated estradiol-17β was inhibited by the P450-aromatase inhibitor formestane. Messenger RNA for P450-aromatase was abundantly expressed both in Sertoli cells in the testis of goats of ages three, four and five months. The expression of P450 mRNA was also localized in Leydig cells. It is concluded that the prepubertal goat has measurable quantities of serum estradiol-17β and that their testis has the capacity to synthesize estrogens during this period.

Serum luteinising hormone, testosterone and total cholesterol levels, libido and testicular histomorphology of male West African Dwarf goats orally or subcutaneously treated with monosodium l-glutamate

This study investigated the effects of administration of monosodium l-glutamate (MSG) on serum luteinising hormone, testosterone and cholesterol levels, libido and testicular histomorphology of male West African Dwarf (WAD) goats. Thirty-two WAD goats (28 males and four females) were used for the study. The 28 males were randomly assigned to seven groups (A, BO, BS, CO, CS, DO and DS) of four goats each. Goats in groups BO, CO, and DO were given MSG orally at doses of 0.25, 0.50 and 1 g/kg body weight respectively for 28 days, while groups BS, CS and DS goats were given MSG subcutaneously at doses of 0.25, 0.50 and 1 g/kg body weight, respectively, for 28 days. Group A goats served as untreated control. The four female goats did not receive MSG, but were used to assess the levels of expression of libido by all the males. Serum luteinising hormone and testosterone were assayed prior to MSG administration (Day 0) and on Days 2, 14 and 28 of MSG administration, while serum cholesterol was assayed on Days 0, 14 and 28 of MSG administration. Libido scores and testicular histomorphology were evaluated on Days 26 and 28 of MSG administration, respectively. Results showed that on Days 14 and 28 of MSG administration the mean serum luteinising hormone, testosterone and cholesterol levels of the treated groups were significantly (P < 0.05) lower than those of the control group. The mean libido scores of all the treated groups were significantly (P < 0.05) lower than that of the control. Sections of the testes of the male WAD goats that received varying doses of MSG orally or subcutaneously showed no obvious lesions; the seminiferous tubules and interstices were normal and comparable to those of the untreated control. It was concluded that MSG administration for up to 14 and 28 days led to a significant lowering of serum luteinising hormone, testosterone and cholesterol, as well as libido scores.

The Tet1 and histone methylation expression pattern in dairy goat testis

DNA methylation and histone methylation are critical for mammalian development. Ten-eleven translocation (Tet1), a key regulator of DNA methylation, has been identified as a key enzyme for the activation of DNA demethylation; histone H3 lysine 9 (H3K9) and 27 (H3K27) methylation repress gene expression. Significant progress on the biological functions of Tet proteins has been made in mice and humans. However, their expression pattern and function in the male germ cells in the dairy goat testis are still unclear. The present study described the expression pattern of Tet1, H3K9, and H3K27 in the dairy goat testis and cultured goat spermatogonia stem cells (gSSCs). The results showed that Tet1 was weakly expressed in the dairy goat's testis compared to other organ tissues. Tet1, 5-hydroxymethylcytosine, H3K9, and H3K27 expressions were positive and dynamically changing during spermatogenesis; however, they showed weak expression in neonate stage in vivo. Tet1 and 5-hydroxymethylcytosine showed low expression in gSSCs in vitro in differentiated cultures. These will provide new perspectives for DNA methylation/demethylation and better regulation of epigenetic modifications in gSSCs.

Dual functions in response to heat stress and spermatogenesis: characterization of expression profile of small heat shock proteins 9 and 10 in goat testis.

Small heat shock proteins 9 and 10 (HSPB9 and HSPB10) are two testis-specific expressed sHsps. The objective of this study was to investigate the mRNA expression profile of HSPB9 and HSPB10 in goat testis among the different seasons, ages, and environmental temperatures. Allocation of the two sHsps was also performed by immunohistochemistry. The results showed that the transcript levels of HSPB9 and HSPB10 were extremely high in the testis (P < 0.01). The relative expression of HSBP9 and HSPB10 in testis showed a tendency to increase with age and then is maintained at the constant level after sexual maturity. HSPB9 and HSPB10 have significantly higher expression in the breeding season (P < 0.05) and hot season (P < 0.01). Both HSPB9 and HSPB10 were found to be upregulated by high-temperature stress in testis (P < 0.05), and the expressions of Hsp70 and Hsp90 were also increased simultaneously (P < 0.01). Immunohistochemistry analysis localized HSPB9 expressed in spermatogonia, spermatocytes, and round spermatids and HSPB10 expressed in the elongate spermatids. In epididymis, strongly staining signal of HSPB10 was detected in pseudostratified columnar epithelium. In conclusion, the two testis-specific sHsps are closely related to male reproduction and heat tolerance. The results could provide valuable data for the further studies on HSPB9 and HSPB10.

A new approach for cryoprotectant-free freezing of goat epididymal spermatozoa.

A cryoprotectant-free method was successfully used for rapid freezing of goat epididymal spermatozoa. Lowering sperm volume may increase the temperature exchange rate and improve the freezing output of spermatozoa. The aim of this study was to compare two different packaging types [0.25 ml French straws (FS) and 96-well immune plate (WIP)] for rapid freezing of goat epididymal spermatozoa. Eleven pairs of the goat testes were transferred to the laboratory; cauda epididymidides were dissected and sliced in TRIS-BSA solution for 15 min and temperature 33-35 °C. Sperm concentration was adjusted to 20 × 10(6) ml(-1), and the suspension was subjected to rapid freezing within FS or WIP. The volume of spermatozoa in WIP method was set at 25 μl. Sperm motility, viability and abnormalities, and sperm DNA integrity were compared between two devices. The results showed similar effectiveness of WIP and FS on post-thaw sperm parameters. In conclusion, for cryoprotectant-free rapid freezing of goat epididymal spermatozoa, it is recommended to use WIP instead of French 0.25 ml straws.

Testis mediated gene transfer: In vitro transfection in goat testis by electroporation

Testis mediated gene transfer (TMGT) is a potential tool for making transgenic mice having more than 90% success rate. However, this method needs further standardization before it can be adapted in other species including livestock. In order to standardize the TMGT in goat, buck testes (n=20) collected from the slaughter house were injected with a vector driving green fluorescent protein (GFP) expression under a cytomegalovirus (CMV) promoter. Then, the testes were subjected to electroporation with predetermined voltage, pulse length, pulse interval and number of pulses. Seminiferous tubules were isolated from the electroporated testis and cultured in-vitro. The expression was checked at regular intervals. Green fluorescence was observed on different days in different samples. It suggests transient integration of the plasmid into the seminiferous tubules. This in-vitro transfection of seminiferous tubule using electroporation will provide valuable baseline information.

Expression profile of Nanos2 gene in dairy goat and its inhibitory effect on Stra8 during meiosis.

Nanos2, an RNA-binding protein, belongs to the Nanos gene-coding family and contains two CCHC zinc-finger motifs. In mouse, it plays a pivotal role in male germ cell development, and self-renewal of spermatogonial stem cells. However, little is known of its expression pattern and functions in dairy goat testis. Immunohistochemistry and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) were used to generate the expression profile of Nanos2 in dairy goat testis. Furthermore, its overexpression effects on male germline stem cells (mGSCs) were studied using qRT-PCR, immunofluorescence, dual-luciferase reporter assay and western blotting. Nanos2 is a conservative gene expressed widely in various tissues, especially in pancreas, and it displays higher expression in adult testes than in other age groups. Overexpression of Nanos2 significantly downregulated meiosis-related genes, including Stra8 and Scp3, which induced inhibition of meiosis. Results from dual-luciferase reporter assay and western blotting indicated that Nanos2 directly downregulated Stra8 in goat GmGSCs. Taken together, these results suggest that Nanos2 plays an important role in spermatogonia and that its overexpression restrained meiosis in the dairy goat.

Morphologic and proliferative characteristics of goat type A spermatogonia in the presence of different sets of growth factors.

The present study by using different growth factors was aimed to develop the best practical culture condition for purification of goat undifferentiated SSCs and their colonization under in vitro and in vivo conditions. The enzymatically isolated SSCs obtained from one month old goat testes were cultured in DMEM plus FCS supplemented with different sets of growth factors (GDNF, LIF, bFGF, and EGF) for 2 weeks. At the end of each week, the morphological characteristics of cells and colonies alongside with purification rate of undifferentiated type A spermatogonia were evaluated by immunocytochemical staining and flow cytometry. The number and size of colonies in treatment groups were significantly (P < 0.01) higher than corresponding values in control group. In immunocytochemical evaluation, the proportion of KIT and PGP9.5 positive cells were significantly (P < 0.001) higher in control and treatment groups, respectively. The culture medium comprising all four growth factors, especially the one supplemented with the higher concentration of GDNF, was superior to the other groups with respect to the population of undifferentiated type A spermatogonia and its propagation in culture system. Additionally, goat SSCs could colonize within the mouse testis following xenotransplantation.

Long-term culture and analysis of cashmere goat Sertoli cells.

Sertoli cells have important functions in the testis for spermatogenesis. Thus, Sertoli cell culture systems have been established in many animals, such as rat, mouse, human, dog, cow, and pig, but a goat culture has not been reported. This study describes the isolation and culture of Sertoli cells from 3- to 4-month-old cashmere goat (Capra hircus) testes. These proliferative cells were expanded for 20 passages and repeatedly cryopreserved in vitro, in contrast to previous study in human, of which maintain steady growth for up to seven passages and only passages 1 to 5 could be refrozen. The microstructure and ultrastructure of the culture were typical of Sertoli cells, bearing irregular nuclei and a cytoplasm that was rich in smooth and rough endoplasmic reticulum, mitochondria, Golgi, lysosomes, lipid drops, and glycogenosomes. By immunofluorescence analysis, the all cells expressed SRY-related HMG box gene 9 (Sox9). Growth curves and 5-bromo-2'-deoxyuridine (BrdU) incorporation were used to analyze the proliferation of the cultured cells. With increasing passage times, the proliferation of the Sertoli cells declined, but the transcription of glial cell-derived neurotrophic factor (GDNF), stem cell factor (SCF), and β1-integrin was constant. By flow cytometry, the cells retained the ability to proliferate after 5 yr of cryopreservation. Thus, cashmere goat Sertoli cells have significant proliferative potential in vitro, expressing germ cell regulatory factors and have important applications in studying Sertoli cell-germ cell interactions, spermatogenesis, reproductive toxicology, and male infertility.

Temporal changes in histomorphology and gene expression in goat testes during postnatal development.

Testicular cell proliferation and differentiation is critical for development of normal testicular function and male reproductive maturity. The objective of the current study was to evaluate histoarchitecture and expression of genes marking specific cells and important functions as well as testosterone production of the developing goat testes. Testes were harvested from Alpine bucks at 0, 2, 4, 6, and 8 mo of age (n = 5/age group). Paired testes weight increased from 2 to 4 (P < 0.001) and 4 to 6 mo (P < 0.01). The greatest increases in seminiferous tubule and lumen diameters and height of the seminiferous epithelium occurred between 2 and 4 mo (P < 0.001). Genes expressed in haploid germ cells (Protamine1 [PRM1], Outer Dense Fiber protein 2 [ODF2], and Stimulated by Retinoic Acid gene 8 [STRA8]) increased dramatically at the same time (P < 0.001). Expression of other genes decreased (P < 0.05) during testicular maturation. These genes included P450 side chain cleavage (CYP11A1), Sex determining region Y-box 9 (SOX9), Insulin-like Growth Factor 1 Receptor (IGF1R), and Heat Shock Protein A8 (HSPA8). The Glutathione S-Transferase A3 (GSTA3) gene, whose product was recently recognized as a primary enzyme involved in isomerization of androstenedione in man and livestock species including goats, sheep, cattle, pigs, and horses, uniquely peaked in expression at 2 mo (P < 0.05). Follicle-Stimulating Hormone Receptor (FSHR) mRNA abundance tended to steadily decrease with age (P = 0.1), while Luteinizing Hormone Receptor (LHCGR) mRNA abundance in testes was not significantly different across the ages. Testosterone content per gram of testicular tissue varied among individuals. However, testosterone content per testis tended to increase at 6 mo (P = 0.06). In conclusion, major changes in cellular structure and gene expression in goat testes were observed at 4 mo of age, when spermatogenesis was initiated. Male goats mature rapidly and represent a good model species for the study of agents that enhance or impair development of testicular functions.

Conservation and function of Dazl in promoting the meiosis of goat male germline stem cells

Dazl (deleted in azoospermia-like) is a conserved gene in mammalian meiosis, which encodes RNA binding protein required for spermatocyte meiosis. Up to date, the expression and function of Dazl in the goat testis are unknown. The objectives of this study were to investigate the expression pattern of Dazl in dairy goat testis and their function in male germline stem cells (mGSCs). The results first revealed that the expression level of Dazl in adult testes was significantly higher than younger and immature goats, and azoospermia and male intersex testis. The dairy goat Dazl is highly conserved analysed by several online and bioinformatics software, respectively. Over-expression of Dazl promoted the expression of meiosis-related genes in dairy goat mGSCs. The expression of Stra8 was up-regulated by over-expression of Dazl analysed by Luciferase reporter assay. Taken together, results suggest the Dazl plays an important role in dairy goat spermatogenesis and that over-expression of Dazl may promote Stra8 expression in dairy goat mGSCs.

Antibody preparation and identification of the Cashmere goat c-kit protein in the testes.

The c-kit protein plays a major role in the regulation of germ cell development. Its expression and distribution in rodent testes have been widely reported. However, research regarding c-kit expression in domestic animals is scarce, and the expression pattern and distribution of c-kit in germ cells have not been clearly defined. In this study, a specific antigenic region for goat c-kit was designed, and a c-kit polyclonal antibody was prepared. This antibody was then applied in a study evaluating c-kit expression in Cashmere goat tissues. A Western blot analysis showed that three forms of c-kit were expressed in goat testes: precursor, mature, and soluble c-kit. Fluorescent immunohistochemical analyses showed that c-kit was primarily expressed in the spermatogonia and spermatocytes of goat testes. These results not only clarify the expression and localization of c-kit in the goat testis, but also accelerate further research regarding the function of c-kit in goat spermatogenesis.

Isolation and phylogenetic analysis of an orf virus from sheep in Makhdoom, India

Orf (contagious ecthyma) is an exanthematic disease caused by a parapoxvirus and occurs primarily in sheep and goats with zoonotic implications. In the present investigation, an orf outbreak in the Muzzaffarnagari sheep flock at the Central Institute for Research on Goats (CIRG), Makhdoom, Mathura, Uttar Pradesh, India, was investigated. Primary goat testes cell culture was used for isolation of the orf virus (ORFV) for the first time. The identity of the virus was confirmed by amplification and sequence analysis of the major envelope glycoprotein (B2L) gene and named ORFV/sheep/India/2012/CIRG. On phylogenetic analysis of B2L protein gene, it clustered with the ORFV strains from China suggesting distinct ORFV strains are circulating in India. On comparison of nucleotide and deduced amino acid sequence analysis (n = 63), a unique 126S residue was observed in ORFV/sheep/India/2012/CIRG. On further sequence analysis (B2L) of different ORFV strains (n = 63), some conserved amino acid residues were identified as host-specific (sheep, human, camel, takin, and musk ox) and have been summarized.

Expression pattern of Ngn3 in dairy goat testis and its function in promoting meiosis by upregulating Stra8

Ngn3 is a typical transcription factor and marker of differentiating spermatogonial stem cells (SSCs) in mouse, belonging to the basic helix-loop-helix (bHLH) family. Its gene is specifically expressed in A type spermatogonia in mouse testis, thus plays a critical role in controlling differentiation of SSCs. However, roles of Ngn3 and its protein in dairy goat testis remain unknown. Testis development and expression patterns of Ngn3 were analysed by immunofluorescence and quantitative reverse transcription-polymerase chain reaction (QRT-PCR) in the dairy goat. Furthermore, effects of its overexpression on male germline stem cells (mGSCs) were evaluated by QRT-PCR, immunofluorescence, luciferase reporter assay and western blotting. Revealed that Ngn3 was expressed more highly during puberty and in the adult than in testis of other ages. Overexpression of Ngn3 promoted expression of meiosis-related gene Stra8 and stem-cell differentiation marker CD117, but suppressed expression of Plzf, a classical marker of SSCs. Furthermore, Ngn3 did not promote expression of Stra8 directly as shown in transcription and translation levels detected by luciferase reporter assay and western blotting. Taken together, these results suggest that Ngn3 plays an important role in spermatogenesis and that overexpression of Ngn3 can promote meiosis in testis of the dairy goat.

Enrichment and characterization of Thy1-positive male germline stem cells (mGSCs) from dairy goat (Capra hircus) testis using magnetic microbeads

In mammalian testis, male germline stem cells (mGSCs) are originated from primordial germ cells and developed into spermatocyte and spermatid. In our previous studies, we had isolated a pluripotent mGSCs from goat testes and tested their pluripotency and differentiation potential in vitro and in vivo, which revealed that the isolated and cultured dairy goat mGSCs maintained the characteristics of mGSCs. However, Thy1, a marker of mGSCs, was not examined in detail. In this study, the dairy goat mGSCs were purified by differential plating followed by magnetic-activated cell sorting (MACS) using Thy1 antibody. The quantitative reverse transcription polymerase chain reaction and immunofluorescence analyses revealed that the transcription and expression of Thy1, CD49f, Plzf, Oct4, Gfra1, and Vasa were higher in Thy1-positive cells when compared with Thy1-negative cells. The detection results of culturing dairy goat mGSCs indicated that the Thy1-positive cells maintained the characteristics of mGSCs, grew relatively faster than Thy1-negative cells, and the percentage of alkaline phosphatase-positive cells and colonies were significantly higher in Thy1-positive mGSCs than Thy1-negative cells. Collectively, these results indicate that THY1 is a marker of undifferentiated spermatogonia in goat testes, the technique of magnetic-activated cell sorting using Thy1 antibody could be an efficient method to enrich mGSCs in goat.