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2011, Number 2

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Rev Mex Med Repro 2011; 3.4 (2)

Calmodulin application as a sperm capacitator

Arellano CMD, Borrego A, Soto CE, Rivas CR

Language: Spanish
References: 18
Page: 86-91
PDF size: 150.82 Kb.


ABSTRACT

Objective: To show the behavior of human sperm exposed to calmodulin (CaM) and other factors, such as 4-(2-hydroxyethyl)-1- piperazine-ethanesulfonic acid (HEPES), adenosine triphosphate (ATP) and voltages.
Material and method: The sample was obtained from three patients with five days of sexual abstinence. We used a Makler chamber (obtained from Invitrogen®), a patch clamp amplifier EPC 7 (HEKA Elektronik) and an analog-digital acquisition of 12 bits (Indec Systems) and 4-(2-hydroxyethyl)-1-piperazine-ethanesulfonic acid (HEPES), adenosine triphosphate (ATP) and calmodulin (CaM), obtained from Sigma-Aldrich. The samples were refrigerated fresh at 4°C for 24 hours. Subsequently, semen samples was performed directly (EBD) using the criteria of the World Health Organization (1999) and taking into account the volume of the sample, the number of sperm contained in each milliliter and the percentage of sperm motility and it was classified as follows: A) rapid progressive movement, B) slow progressive motion, C) in situ, and D) without movement.
Results: Samples movement lost after 24 hours under refrigeration, were measured 42 X 106 sperm without movement. One milliliter of HEPES solution was used and recovered sperm movement (B + C) in 80%. The use of CaM (1 mg/uL) and ATP (1 mM) also activated the sperm, as 90 to 100% of these recovered movement (A + B). In the control group there was lost of movement at the hour 8, with CaM motion was lost approximately in 20 min, and molecules CaM and ATP are apparently associated because both molecules are involved in maintaining the balance of Ca2+, which conferred more sperm movement time (approximately 20 hours). In the control group (treatment 1) sperm lost movement because they remained for 24 hours at 4°C. With HEPES (treatment 2) sperm movement increased. With the use of CaM-ATP (treatment 3) sperm movement was superior to treatment 1 (control group) and treatment 2 (with HEPES).
Conclusion: With calmodulin (CaM) the motion is lost after 20 min. With the use of HEPES sperm movement increased by phosphates in it. CaM and ATP showed an association, which gave him more time progressive movement (A + B) sperm, and when the sperm were submitted to different electrical stimuli, sperm movement down when higher electrical stimulation is applied, probably by alteration of voltage-dependent proteins that are important for sperm capacitation. The opening of ATP- and CaM-dependent Ca2+ channels of the plasma membrane probably promotes sperm capacitation. With the results of this work calmodulin (CaM) can be established as a new technique of assisted reproduction in artificial insemination in humans.


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