Table 2.
Qualitative Analysis and Grading of p21CIP1/Waf1 Immunohistochemistry.a
| Groups | Stages | Spermatogonia | Primary spermatocytes | MF/SS | Round spermatids | Elongating spermatids | Sertoli cells |
|---|---|---|---|---|---|---|---|
| Control | I-VI | Spermatogonia − | Pachytene − | NA | Steps 1-6 + | Steps 15-18 − | Sertoli cells − |
| VII-IX | Spermatogonia − | Pachytene − Preleptotene − Leptotene − | NA | Steps 7-9 + | Step 19 − | Sertoli cells − | |
| X-XIII | Spermatogonia − | Leptotene – Zygoteine – Pachytene + | NA | NA | Steps 10-13 +++ | Sertoli cells − | |
| XIV | Spermatogonia − | Pachytene − | MF −/+ SS −/+ | NA | Step 14 − | Sertoli cells − | |
| DM1 | I-VI | Spermatogonia − | Pachytene −/+ | NA | Steps 1-6 +++ | Steps 15-18 − | Sertoli cells − |
| VII-IX | Spermatogonia − | Pachytene −/+ Preleptotene −/+ Leptotene −/+ | NA | Steps 7-9 +++ | Steps 19 − | Sertoli cells − | |
| X-XIII | Spermatogonia − | Leptotene −/+ Zygotene −/+ Pachytene −/+ | NA | NA | Steps 10-13 ++ | Sertoli cells − | |
| XIV | Spermatogonia − | Pachytene − | MF +++ SS +++ | NA | Step 14 − | Sertoli cells − | |
| DM3 | I-VI | Spermatogonia − | Pachytene +++ | NA | Steps 1-6 +++ | Steps 14-18 − | Sertoli cells − |
| VII-IX | Spermatogonia − | Pachytene ++++ Preleptotene ++++ Leptotene ++++ | Na | Steps 7-9 ++++ | Step 19 − | Sertoli cells − | |
| X-XIII | Spermatogonia − | Leptotene ++++ Zygotene ++++ Pachytene ++++ | NA | NA | Steps 10-13 +++++ | Sertoli cells − | |
| XIV | Spermatogonia − | Pachytene − | MF +++ SS +++ | NA | Step 14 +++ | Sertoli cells − |
Abbreviations: I-XIV, seminiferous epithelial stages; DM1, 1-month-long diabetic group; DM3, 3-month-long diabetic group; MF, meiotic figures; NA, not applicable as the particular feature is not seen in that tubular stage; p21CIP1/Waf1, protein 21cyclin-dependent kinase interacting protein 1/wild-type p53-activated factor 1; SS, secondary spermatocytes; steps 1-19, different steps of spermiogenesis.
a The grading of immunohistochemical labeling of the protein was done in a blinded manner (N = 6). The protein was localized predominantly in cytoplasm, but also in perinuclear regions of testicular cells. Negative or equivocal labeling was graded as “−,” mild labeling as “+,” moderate labeling as “++,” intense labeling as “+++,” more intense labeling as “++++,” and highly intense labeling as “+++++.”