ABSTRACT
Pathogenesis of Bothrops envenomations is complex and despite numerous studies on the effects of this snake venom on various biological systems, relatively little is known about such effects on the male reproductive system. In the present study, the toxicological outcomes of the low molecular weight fraction (LMWF) of B. jararaca snake venom - containing a range of bioactive peptides - were investigated on the dynamics and structure of the seminiferous epithelium and 15P-1 Sertoli cells viability. Methods: LMWF (5 µg/dose per testis) venom was administered in male Swiss mice by intratesticular (i.t.) injection. Seven days after this procedure, the testes were collected for morphological and morphometric evaluation, distribution of claudin-1 in the seminiferous epithelium by immunohistochemical analyses of testes, and the nitric oxide (NO) levels were evaluated in the total extract of the testis protein. In addition, the toxicological effects of LMWF and crude venom (CV) were analyzed on the 15P-1 Sertoli cell culture. Results: LMWF induced changes in the structure and function of the seminiferous epithelium without altering claudin-1 distribution. LMWF effects were characterized especially by lost cells in the adluminal compartment of epithelium (spermatocytes in pachytene, preleptotene spermatocytes, zygotene spermatocytes, and round spermatid) and different stages of the seminiferous epithelium cycle. LMWF also increased the NO levels in the total extract of the testis protein and was not cytotoxic in concentrations and time tested in the present study. However, CV showed cytotoxicity at 10 μg/mL from 6 to 48 h of treatment. Conclusions: The major finding of the present study was that the LMWF inhibited spermatozoa production; principally in the spermiogenesis stage without altering claudin-1 distribution in the basal compartment. Moreover, NO increased by LMWF induce open of complexes junctions and release the germ cells of the adluminal compartment to the seminiferous tubule.(AU)
Subject(s)
Animals , Male , Peptides , Seminiferous Epithelium , Snake Venoms , Spermatogenesis , Bothrops , Biological ProductsABSTRACT
Objective To investigate the effect of long term low-dose manganese exposure on testicular spermatogenic cell mitochondria morphology and apoptosis of male offspring rat.Methods Thirty-two healthy female SD rats were divided into the control group,low,middle and high dose groups.2,4 and 8 mg/kg MnCl2 · 4H2 O or normal saline was intraperitoneally injected for 8 weeks(once daily,5 d/week).The manganese exposure continued during the pregnant period and lactation period.Eight 12-week-old offspring male rats were killed in each group,the structure of seminiferous tubules and mitochondria morphology in spermatogenic cells were observed.The expression of Opa1,Drp1 and Caspase9,and the apoptosis of spermatogenic cells were detected.Results With the increase of administered-MnCl2 dosage,the number of spermatogenic cell layers decreased,spermatogenic cells arranged in disorder,the number reduced,etc;the mitochondria separation and swelling of spermatogenic cells were found in the middle and high dose groups;the expression of Opa1 was gradually decreased in the middle and high dose groups(P＜0.05,P＜0.01),while the expression of Drp1 and Caspase9 was gradually increased with the manganese dose increase(P＜0.05,P＜0.01);the apoptotic index of spermatogenic cells in the manganese exposure group was significantly increased with the increase of administeredMnCl2 dosage(P＜0.05).Conclusion Long term low dose of manganese exposure could regulate the expression of Opa1/Drp1 gene and affect the function of mitochondria,leads to apoptosis of spermatogenic cells in male offspring rat.
ABSTRACT
Background Considering the similarity between the testis-specific isoform of angiotensin-converting enzyme and the C-terminal catalytic domain of somatic ACE as well as the structural and functional variability of its natural inhibitors, known as bradykinin-potentiating peptides (BPPs), the effects of different synthetic peptides, BPP-10c (<ENWPHQIPP), BPP-11e (<EARPPHPPIPP), BPP-AP (<EARPPHPPIPPAP) and captopril were evaluated in the seminiferous epithelium of male mice.Methods The adult animals received either one of the synthetic peptides or captopril (120 nmol/dose per testis) via injection into the testicular parenchyma. After seven days, the mice were sacrificed, and the testes were collected for histopathological evaluation.Results BPP-10c and BPP-AP showed an intense disruption of the epithelium, presence of atypical multinucleated cells in the lumen and high degree of seminiferous tubule degeneration, especially in BPP-AP-treated animals. In addition, both synthetic peptides led to a significant reduction in the number of spermatocytes and round spermatids in stages I, V and VII/VIII of the seminiferous cycle, thickness of the seminiferous epithelium and diameter of the seminiferous tubule lumen. Interestingly, no morphological or morphometric alterations were observed in animals treated with captopril or BPP-11e.Conclusions The major finding of the present study was that the demonstrated effects of BPP-10c and BPP-AP on the seminiferous epithelium are dependent on their primary structure and cannot be extrapolated to other BPPs.(AU)
Subject(s)
Animals , Mice , Seminiferous Epithelium , Snake Venoms , Angiotensin-Converting Enzyme Inhibitors , Bothrops , Protein IsoformsABSTRACT
Background Considering the similarity between the testis-specific isoform of angiotensin-converting enzyme and the C-terminal catalytic domain of somatic ACE as well as the structural and functional variability of its natural inhibitors, known as bradykinin-potentiating peptides (BPPs), the effects of different synthetic peptides, BPP-10c ( ENWPHQIPP), BPP-11e ( EARPPHPPIPP), BPP-AP ( EARPPHPPIPPAP) and captopril were evaluated in the seminiferous epithelium of male mice.Methods The adult animals received either one of the synthetic peptides or captopril (120 nmol/dose per testis) via injection into the testicular parenchyma. After seven days, the mice were sacrificed, and the testes were collected for histopathological evaluation.Results BPP-10c and BPP-AP showed an intense disruption of the epithelium, presence of atypical multinucleated cells in the lumen and high degree of seminiferous tubule degeneration, especially in BPP-AP-treated animals. In addition, both synthetic peptides led to a significant reduction in the number of spermatocytes and round spermatids in stages I, V and VII/VIII of the seminiferous cycle, thickness of the seminiferous epithelium and diameter of the seminiferous tubule lumen. Interestingly, no morphological or morphometric alterations were observed in animals treated with captopril or BPP-11e.Conclusions The major finding of the present study was that the demonstrated effects of BPP-10c and BPP-AP on the seminiferous epithelium are dependent on their primary structure and cannot be extrapolated to other BPPs.
Subject(s)
Male , Animals , Mice , Angiotensins , Bothrops , Seminiferous Epithelium , Enzyme Inhibitors , Crotalid VenomsABSTRACT
Background The testis-specific isoform of angiotensin-converting enzyme (tACE) is exclusively expressed in germ cells during spermatogenesis. Although the exact role of tACE in male fertility is unknown, it clearly plays a critical function in spermatogenesis. The dipeptidase domain of tACE is identical to the C-terminal catalytic domain of somatic ACE (sACE). Bradykinin potentiating peptides (BPPs) from snake venoms are the first natural sACE inhibitors described and their structure-activity relationship studies were the basis for the development of antihypertensive drugs such as captopril. In recent years, it has been showed that a number of BPPs - including BPP-10c - are able to distinguish between the N- and C-active sites of sACE, what is not applicable to captopril. Considering the similarity between tACE and sACE (and since BPPs are able to distinguish between the two active sites of sACE), the effects of the BPP-10c and captopril on the structure and function of the seminiferous epithelium were characterized in the present study. BPP-10c and captopril were administered in male Swiss mice by intraperitoneal injection (4.7 μmol/kg for 15 days) and histological sections of testes were analyzed. Classification of seminiferous tubules and stage analysis were carried out for quantitative evaluation of germ cells of the seminiferous epithelium. The blood-testis barrier (BTB) permeability and distribution of claudin-1 in the seminiferous epithelium were analyzed by hypertonic fixative method and immunohistochemical analyses of testes, respectively. Results The morphology of seminiferous tubules from animals treated with BPP-10c showed an intense disruption of the epithelium, presence of atypical multinucleated cells in the lumen and degenerated germ cells in the adluminal compartment. BPP-10c led to an increase in the number of round spermatids and total support capacity of Sertoli cell in stages I, V, VII/VIII of the seminiferous epithelium cycle, without affecting BTB permeability and the distribution of claudin-1 in the seminiferous epithelium. Interestingly, no morphological or morphometric alterations were observed in animals treated with captopril. Conclusions The major finding of the present study was that BPP-10c, and not captopril, modifies spermatogenesis by causing hyperplasia of round spermatids in stages I, V, and VII/VIII of the spermatogenic cycle.(AU)
Subject(s)
Animals , Peptides , Seminiferous Epithelium , Seminiferous Tubules , Snake Venoms , Bradykinin , Bothrops/anatomy & histologyABSTRACT
Background The testis-specific isoform of angiotensin-converting enzyme (tACE) is exclusively expressed in germ cells during spermatogenesis. Although the exact role of tACE in male fertility is unknown, it clearly plays a critical function in spermatogenesis. The dipeptidase domain of tACE is identical to the C-terminal catalytic domain of somatic ACE (sACE). Bradykinin potentiating peptides (BPPs) from snake venoms are the first natural sACE inhibitors described and their structure-activity relationship studies were the basis for the development of antihypertensive drugs such as captopril. In recent years, it has been showed that a number of BPPs - including BPP-10c - are able to distinguish between the N- and C-active sites of sACE, what is not applicable to captopril. Considering the similarity between tACE and sACE (and since BPPs are able to distinguish between the two active sites of sACE), the effects of the BPP-10c and captopril on the structure and function of the seminiferous epithelium were characterized in the present study. BPP-10c and captopril were administered in male Swiss mice by intraperitoneal injection (4.7 mol/kg for 15 days) and histological sections of testes were analyzed. Classification of seminiferous tubules and stage analysis were carried out for quantitative evaluation of germ cells of the seminiferous epithelium. The blood-testis barrier (BTB) permeability and distribution of claudin-1 in the seminiferous epithelium were analyzed by hypertonic fixative method and immunohistochemical analyses of testes, respectively. Results The morphology of seminiferous tubules from animals treated with BPP-10c showed an intense disruption of the epithelium, presence of atypical multinucleated cells in the lumen and degenerated germ cells in the adluminal compartment. BPP-10c led to an increase in the number of round spermatids and total support capacity of Sertoli cell in stages I, V, VII/VIII of the seminiferous epithelium cycle, without affecting BTB permeability and the distribution of claudin-1 in the seminiferous epithelium. Interestingly, no morphological or morphometric alterations were observed in animals treated with captopril. Conclusions The major finding of the present study was that BPP-10c, and not captopril, modifies spermatogenesis by causing hyperplasia of round spermatids in stages I, V, and VII/VIII of the spermatogenic cycle.
ABSTRACT
Estudaram-se os efeitos da geleia real sobre a espermatogênese de coelhos tratados com diferentes concentrações de geleia real. Os tratamentos foram formados por três grupos: grupo-controle; grupo que recebeu 0,5mg/dia de geleia real; e grupo que recebeu 1,0mg/dia de geleia real. O estudo envolveu a morfometria testicular. Não houve diferença entre os tratamentos quanto aos pesos corporal (T1=3,20±0,19kg, T2=2,96±0,30kg e T3=3,21±0,37kg) e gonadal (T1=2,36±0,33g, T2=2,53±0,33g e T3=2,64±0,39g) e quanto aos índices gonadossomático (T1=0,15±0,02%, T2=0,17±0,03% e T3=0,16±0,02%) e tubulossomático (T1=0,06±0,01%; T2=0,07±0,01% e T3=0,06±0,01%). O diâmetro médio dos túbulos seminíferos (T1=225,95±13,27µm, T2=239,68±21,50µm e T3=231,57±15,94µm), a altura do epitélio seminífero (T1=66,05±5,37µm, T2= 73,47±9,11µm e T3=63,34±4,79µm) e o comprimento de túbulos seminíferos por testículo (T1=46,63±13,44m, T2=43,58±12,17m e T3=46,96±9,54m) e por grama de testículo (T1=19,50±2,68m, T2=17,12±3,91m e T3=17,78±1,98m) não diferiram entre tratamentos. Conclui-se que a suplementação com geleia real, nas doses utilizadas, não altera as características testiculares avaliadas.
This study aimed to investigate the effects of royal jelly on spermatogenesis in rabbits treated with different concentrations of RJ (Control; 0,5mg/day; and 1,0mg/day) using testicular morphometry. There was no significant difference between the body weight (T1= 3.20±0.19kg; T2= 2.96±0.30kg; T3=3.21±0.37kg) and gonadal weight (T1= 2.36±0.33g; T2= 2.53±0.33g; T3= 2.64±0.39g), gonadossomatic index (T1= 0.15±0.02%; T2= 0.17±0.03%. T3= 0.16±0.02%) and tubulossomatic index (T1= 0.06±0.01%; T2= 0.07±0.01%. T3= 0.06±0.01%) between treatments, showing that the percentage of body mass, and the percentage of seminiferous tubules allocated in testis were similar in the 3 experimental groups. Similarly, the mean diameter of the seminiferous tubules (T1= 225.95±13.27µm; T2=239.68±21.50µm; T3= 231.57±15,94µm), the height of the seminiferous epithelium (T1=66,05±5,37µm; T2=73.47±9.11µm; T3=63.34±4.79 µm) and length of seminiferous tubule for testis (T1=46.63±13.44m; T2=43.58±12.17m; T3=46.96±9.54m) and per gram of testis (T1=19.50±2.68m; T2=17.12±3.91m; T3=17.78±1.98m) did not differ statistically. It was concluded that supplementation with royal jelly, at the doses used, did not alter the testicular parameters evaluated here.
Subject(s)
Animals , Rabbits , Spermatogenesis/physiology , Seminiferous Epithelium , Testis/anatomy & histology , Seminiferous Tubules/metabolism , Bees , Microscopy/veterinaryABSTRACT
The species Heteropterys aphrodisiaca is commonly used as a stimulant by popular medicine in the Cerrado, a savanna-like biome, Brazil. Recent studies have proved its protective effects on testes of animals submitted to treatment using Cyclosporine A, as well as its stimulus effect in increasing testosterone secretion. Therefore, the present study was designed to analyze whether the association of the plant infusion and endurance exercise could potentiate the stimulating effect. The animals were separated into 4 groups: two control (sedentary and trained) receiving water and two treated (sedentary and trained) receiving the plant infusion daily (104mg/day). The proportion of the seminiferous tubule compartment and interstitium was analyzed. Within the seminiferous epithelium, the number of Sertoli and germ cells were counted in order to evaluate whether the treatment would alter the spermatogenic dynamics, analyzing: the spermatogenic yield, the mitotic and meiotic indexes, the total number of germ cells and the Sertoli cell support capacity. Trained and treated animals showed increased spermatogenic yield and spermatogonia mitosis, and no significant differences in apoptotic indexes. Despite the results showing the same pattern regarding yield and mitotic index, the meiotic index was higher in the sedentary/treated group. Therefore, the H. aphrodisiaca infusion increased both the testosterone production and the spermatogonia mitosis, thus increasing the spermatogenic yield.
Subject(s)
Animals , Male , Rats , Germ Cells/physiology , Malpighiaceae/chemistry , Physical Conditioning, Animal/physiology , Physical Endurance/drug effects , Plant Extracts/pharmacology , Spermatogenesis/drug effects , Testis/drug effects , Apoptosis/physiology , Physical Endurance/physiology , Plant Extracts/administration & dosage , Rats, Wistar , Testis/pathology , Testosterone/physiologyABSTRACT
A cutia (Dasyprocta aguti) é um roedor silvestre encontrado amplamente na região Nordeste do Brasil. É uma espécie muito utilizada pela população humana de baixa renda como fonte alternativa de proteína na alimentação. Foram utilizadas 31 cutias, machos, provenientes da Universidade Federal do Piauí, Estado do Piauí e da Escola Superior de Agricultura de Mossoró Estado do Rio Grande do Norte. Os animais foram divididos em grupos etários desde o nascimento até os 14 meses de idade. O diâmetro nuclear médio foi obtido pela medida de 10 núcleos do tipo celular estudado em cada testículo, no estágio 1 do ciclo do epitélio seminífero. Nos animais que não apresentaram o epitélio organizado em estágios bem definidos em virtude da idade, foram feitas medidas em secções transversais escolhidas somente pelo contorno circular. O início da assincronia do processo espermatogênico foi observado a partir dos seis meses de idade. A puberdade, na cutia Dasyprocta aguti, foi definitivamente estabelecida a partir dos nove meses de idade, pois estavam presentes todos os tipos celulares e espermatozóides liberados no lume tubular em grande parte do parênquima testicular.
The Agouti (Dasyprocta aguti) is a wild rodent that, in the Northeast region of Brazil, is a species that is very used by the low-income population as an alternative protein source for human feeding. Thirty-one male Agouti were used, coming from the Universidade Federal do Piauí (UFPI) - Piauí State, and from Escola Superior de Agricultura de Mossoró (ESAM) - Rio Grande do Norte State - Brazil. The animals were divided in to age groups, from the birth to fourteen months old. The average diameter average of the nucleus was obtained by measuring 10 nuclei of the studied cellular type, in each testicle that was in the stage I of the seminiferous epithelium cycle. This was performed in animals that did not present an organized epithelium in well-defined periods due to the age, and measurements were made in transversal sections chosen only for the circular contour. The beginning of the asynchronism of the spermatogenic process was observed since the six months of age. The puberty of the Agouti Dasyprocta aguti was definitely established at nine months of age, because all the cellular types and free spermatozoa in the tubular lumen were present in a large extent of the testicular parenchyma.
ABSTRACT
Objective To study the effects of experimental varicocele on the apoptosis of spermatogenic cell in adolescent rats.Methods Experimental varicocele models were created by partial ligation of left renal vein in Sprague-Dawley(SD) male rats.The apoptosis of spermatogenic cells was detected by in situ terminal deoxylnucleotidyl transferase mediated-dUTP nick end labeling(TUNEL) technique.Results The incidence of apoptosis was increased significantly in experimental group than that in control group(P
ABSTRACT
Number of germ cells in the seminiferous epithelium was analyzed quantitatively in testicular biopsy specimens of 23 patients without ductal obstruction and of 4 patients with ductal obstruction. Roth number of mature spermatids within each cross-section of seminiferous tubule and number of atrophic tubule were counted in biopsy specimens. Results were expressed as cell number of mature spermatids per seminiferous tubule and percentage of atrophic tubules. A significant correlation was demonstrated between sperm density and mature spermatid counts. Patients with sperm counts of less than 40 x l0(6)/ml had mature spermatids counts of less than 25 per seminiferous tubule. Coefficients of correlation between mature spermatid count and percentage of atrophic tubules were higher than those of correlation between sperm counts and percentage of atrophic tubules. In asoospermrc patients with epididymal obstruction, sperm count after corrective surgery could be predicted correctly by this quantitative analysis technique of testicular biopsy specimens and partial obstruction of anastomotic site of seminal tract could be proved in oligozoospermic patients after corrective surgery.
Subject(s)
Humans , Biopsy , Cell Count , Germ Cells , Seminiferous Epithelium , Seminiferous Tubules , Sperm Count , Spermatids , Spermatozoa , TestisABSTRACT
To investigate the function of microtubules (MTs) in Sertoli cell, colchicine, a microtubule disrupting agents, was injected into the dorsum of the male SD rat. The seminiferous tubules and Sertoli cell of the rat were examined by light and electron microsopy. 5-6 hours after the colchicine injection the following results were revealed (1) The Sertoli cell of the rat testis showed prominent morphological changes. The processes of the cell retracted, the MTs disappeared in the cytoplasm and the distribution of mitochondria and smooth endoplasmic reticulum became irregular. (2) Many widened intercellulor spaces occured in the seminiferous epithelium; and accordingly the arrangement of the germ cells became deranged and the elongated spermatids moved form their original deep position to the surface of the epithelium. (3) There was no evident changes in the basal region of the seminiferous epithilium. These results confirmed that MTs in Sertoli cell are essential to the maintenance of the normal shape and cellular organelle arrangement in the cell as well as the architecture of seminiferous epithelium. They may also play an important role in the movement of the elongated spermatids.