| The diabetes mellitus(DM)and thyroid dysfunction(TD)are found to subsist in chorus.These endocrinal conditions affect not only human population,but also have been reported in many animal species worldwide.Insulin and thyroid hormones are known to affect directly the cellular activities of the testis and epididymis;consequently these play a pivotal role in the regulation of healthy reproduction.The blood-testis barrier(BTB)is one the tightest hemato-tissue barrier of the body and the Claudins-11 protein plays an important role in maintaining the integrity of the tight junction at BTB.But the literature is silent about the expression and immuno-localization of Claudin-11 in the testis during DM and TD diseases.Furthermore,the effects of these concurrent metabolic dysfunctions on different systems of the body have been investigated and discussed only as retrospective studies on the bases of clinical case records in humans,but the data was found lacking in the context of research trials to examine such a grave syndrome and its effects on the male fertility.Thus the present study was conducted to investigate the effects of experimental contemporaneous STZ-diabetes mellitus,hyperthyroidism and hypothyroidism on the weights of body,testes and epididymides,endocrine hormonal profiles,histoarchitecture of testis and epididymis,spermatogenesis and immune-localization of claudin-11 in the seminiferous tubules of pre-to post-pubertal male mice.1.Effects of contemporaneous diabetes mellitus cum hyperthyroidism on the fertility of male mice and immuno-localization of Claudin-11 inside the seminiferous tubulesIn total,32 ICR strain mice were equally distributed into four groups:control(C),diabetic(D),diabetic-plus-hyperthyroid(DH),and hyperthyroid(H).Mice allocated for diabetes,received a single intraperitoneal injection of streptozotocin(STZ)at a dose of 200 mg/kg body weight for the induction of type-1 DM.At the onset of diabetes,one group of the mice was concomitantly injected levothyroxine(LT4)at the dose rate of 0.3 mg/kg body weight and an other set of the animals received the same treatment independently,on daily basis.The body weight along with the weights of testes and epididymides were reduced markedly in the D and DH mice versus control.Higher trends of blood glucose levels were seen in the DH group in comparison to the euthyroid diabetic mice.Our experimental results suggested that the thyroid hormones could exert a transient effect on blood glucose homeostasis by altering the serum blood glucose level in the diabetic patients.The histo-metric analysis of the testes showed increased luminal size of the seminiferous tubules,along with decreased epithelial height and atrophic changes in the germinal stem cells of DH and H mice.Caput epididymis(CpE)of the DH mice showed extensive compaction of principal cells,loss of stereocilia,lipid vacuolization,and inflammatory infiltrations;however,the cauda epididymis(CdE)of the same group of mice exhibited profoundly damaged tubular integrity,packed clear cells,exfoliated cells,and round spermatids.Hyperthyroidism elevated the serum testosterone levels in the H and DH mice and produced critical damages to the histoarchitecture of the epididymis,which were presumed because of the deficiency of gonadotropins in these animals.The control animals of this study,expressed robust staining of the claudin-11 by forming regular zigzag border like structure in between Sertoli cells.In addition,the hyperthyroidism showed upregulatory effects on expression of the claudin-11,by showing positive strong expression of claudin-11 in between Sertoli cells in H(72%),followed by DH(24%)versus controls.In the light of our experimental findings,the mechanisms involved in such alterations of expression patterns of claudin-11 needs to be validated by further experiments with longer study periods.2.Effects of concomitant diabetes mellitus plus hypothyroidism on the male fertility and immuno-localization of Claudin-11 inside the seminiferous tubules of miceTotal,32 ICR strain mice were equally distributed into four equal groups;control(C),diabetic(D),diabetic-plus-hypothyroid(Dh)and hypothyroid(h).Mice allocated for the induction of diabetes mellitus,received a single intraperitoneal injection of STZ at a dose of 200 mg/kg body weight.At the onset of diabetes,one group of mice were concomitantly rendered hypothyroid by adding Methimazole(MMI)0.05%plus potassium perchlorate(KCLO4)0.5%in their drinking water,and an other set of the animals received the same treatment independently.This study observed that,the experimental hypothyroidism and DM can significantly decrease the body,testes and epididymal weights in the D,Dh and h animals.After three weeks of induction of the diabetes and hypothyroidism,the syndrome group(Dh)of mice showed lower blood glucose levels in comparison to euthyroid diabetic animals.The concentration of serum testosterone and thyroidal hormones(T4 and T3)were found diminished in all treated animals.The histo-pathological observations of the testicular and epididymal sections of mice revealed that STZ-diabetes and hypothyroidism posed detrimental effects of various degrees in the seminiferous tubules(St)and ductus epididymis.Irregularly outlined St with increased size of lumina were noticed in the testes of D and h animals.The epididymides of syndrome group were found gravely damaged,and increased interstitial stroma,inflammatory infiltrations,hyperplasia of the principal and clear cells,and exfoliated germ cells were prominent features,observed with variable degrees in the CpE and CdE of Dh,D and h animals.The control animals of this study,expressed the robust staining of Claudin-11 by forming regular zigzag border like structure in between Sertoli cells,but the amount and pattern of this protein localization was seen impaired in treated mice.The animals of syndrome group exhibited downregulated(73%)expressions of Claudin-11,followed by h(46%)and D(37%),compared with controls.Furthermore,punctuate and dislocated expression of this protein was seen on the germinal cells in treated animals.This type of down-regulation of Claudin-11 indicates the loss of functional integrity of the BTB,which suggests the dysfunction of the protein,but not the failure of its expression.3.Effects of concurrent hyperthyroidism and diabetes mellitus on the male fertility and spermatogenesis in pre-and post-pubertal miceFollowing parturition of 16 primiparae ICR pregnant female mice,forty-eight male pups were randomly selected and divided into four equal(n=12/group)groups;C,D,DH and H.The pups of D and DH were induced diabetes by three consecutive STZ injections(40mg/kg)on post-natal day 3,4 and 8.The pups of DH and H were rendered hyperthyroid through breastfeeding,by injecting their dams with LT4(0.3 mg/kg body weight)on daily basis.After weaning(24d)half of the pups were sampled and remaining half continued to get the same LT4 treatment individually till 56 days.During this experiment,eighty percent neonatal mice of the syndrome group exhibited no hairs on the body(alopecia).The body and testicular weights were severely diminished in the syndrome group at both study periods,although these weights were raised to normal healthy levels during second phase of the study.Our experiment revealed diminished fT4 and fT3 levels in the serum of both pre-and post-pubertal diabetic mice.However,in contrast these values were found profoundly elevated in LT4 treated animals.Histo-morphometric observations of this study,determined increased luminal diameter of the St in all treated animals.The shrinkage of germinal epithelium was seen more severe in DH and D group animals at both study periods.During pre-pubertal study,the most of the St of the syndrome group of animals have shown undifferentiated and sloughed epithelial cells inside the lumen.While,markedly wider lumens and vacuolization of the germinal epithelium were noticed as prominent histo-pathologies in the St of the D and H subjects respectively.The diameter and luminal size of CdE in the DH animals significantly diminished at both pre-and post-pubertal studies.Furthermore,in comparison to control,the epithelial height of CpE significantly increased by 43%and 18%in adult mice of the DH and H groups respectively.Similarly,epithelial height of CdE in post-pubertal animals was observed severely elevated in the DH and H(91%)and D(27%)mice.This study has determined that STZ-diabetes causes intense diminution of sperm count and progressive motility in the pubertal mice versus all other experimental subjects.4.Effects of coexisting Diabetes Mellitus and Hypothyroidism on the testicular and epididymal histo-architecture and spermatogenesis of neonatal to adult miceFollowing parturition of 16 primiparae ICR pregnant female mice,forty-eight male pups were randomly selected and divided into four equal(n=12/group)groups;C,D,Dh and h.The pups of D and Dh mice were induced diabetes by three consecutive STZ injections(40mg/kg)on post-natal day 3,4 and 8.The pups of Dh and h were rendered hypothyroid through breastfeeding by adding MMI plus KClO4 in the drinking water of their dams.After weaning(24d)half of the pups,were sampled and remaining half continued to get the same hypothyroid treatment individually,till 56 days.The weights of body,testes and epididymides,hormonal values and morpho-metric measurements together with histo-pathologic features were observed intensely impaired in Dh followed by h and D subjects versus controls at both study periods.The total number of spermatozoa and progressive motility was critically reduced in the Dh and h,followed by D animals. |
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