Defective expression of Galpha12 in the testes of azoospermia patients and in the spermatozoa with low motility.

Antibody to the Galpha12-subunit of guanine nucleotide regulatory proteins was used to determine whether the Galpha12 is present in adult human spermatogenic cells and to determine its role in dyszoospermia. Immunoblots from testes and spermatozoa demonstrated the presence of Galpha12 in the samples. Immunohistochemical analyses of testes found that Galpha12 was expressed in the cytoplasm of Leydig cells and was expressed in spermatids from the elongating Sb phase to mature sperm. Indirect immunofluorescence of human spermatozoa revealed the presence of Galpha12 in the neck region and the midpiece of the sperm. Galpha12 in spermatids and spermatozoa partially co-localized with F-actin and alpha-tubulin. Immunohistochemical analyses of tissues from three patients with non-obstructive azoospermia showed abnormal expression of Galpha12 in more than 45% of spermatids. Furthermore, Western blots and indirect immunofluorescence found defective expression of Galpha12 in low-motility spermatozoa with midpieces that were bent on themselves. Therefore, it suggests that Galpha12 plays a role in polarity and tail formation as spermatids mature. Furthermore, Galpha12 may be a candidate protein responsible for azoospermia caused by spermatogenic disturbance or midpiece deformities.