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Jackson C Kirkman-Brown

Publications and source records attributed to Jackson C Kirkman-Brown.

5 recordsLinked to original sources
PubMed2006

Kinetics of the progesterone-induced acrosome reaction and its relation to intracellular calcium responses in individual human spermatozoa.

Progesterone at 3 microM triggers a biphasic (transient and sustained) increase in intracellular calcium ([Ca(2+)](i)) in human sperm, which is believed to be a prerequisite for progesterone-induced acrosome reaction (AR). As very little is known about how AR occurrence, latency, and completion relate to the characteristics of the progesterone-induced [Ca(2+)](i) signal, we examined these events using fluorescence microscopy of individual living human sperm. Direct assessment of acrosomal status after calcium imaging showed no differences in kinetics or amplitude of the preceding progesterone-induced calcium responses in acrosome-reacted and acrosome-intact cells, which indicates that the amplitude of the [Ca(2+)](i) signal is not the critical determinant of AR. Chelation of extracellular calcium to arrest AR at varying times after progesterone stimulation revealed that maximal AR occurred immediately following progesterone stimulation, during the initial transient calcium influx rather than during the sustained calcium response. Attempts to follow acrosomal dispersal in real-time by staining with the acidic organelle probes LysoTracker DND-99 and dapoxyl (2-aminoethyl) sulphonamide (DAES) proved inconclusive due to heterogeneous labeling of the cell population. Surprisingly, the dye was often not confined to the acrosome but stained the whole sperm head, which suggests that only a subpopulation of human sperm cells contains a sufficiently acidic acrosome.

Acids↗
PubMed2004

Slow calcium oscillations in human spermatozoa.

We have used single-cell imaging to investigate intracellular Ca2+ signalling in human spermatozoa stimulated with progesterone (3 microM). In approx. 9% of cells progesterone caused the activation of slow repetitive [Ca2+]i (intracellular Ca2+ concentration) oscillations, with a period of 1-4 min, which persisted for the duration of recording (20-30 min). Pretreatment with nifedipine, which blocks T- and L-type voltage-operated Ca2+ channels in spermatogenic cells, did not modify the characteristics of the oscillations, but reduced the proportion of cells in which they were observed. Stimulation with Bay K 8644 or FPL64176 induced [Ca2+]i oscillations in 5-10% of cells, but their frequency was low (period, 4-5 min). Application of valinomycin (1 microM) to clamp membrane potential at E(K) (equilibrium potential for potassium) did not modify activity in oscillating cells, showing that plasma membrane potential and activation of voltage-operated conductances are not involved in the mechanism by which sperm [Ca2+]i oscillations are generated.

Calcium Channel Agonists↗
PubMed2003

Encoding of progesterone stimulus intensity by intracellular [Ca2+] ([Ca2+]i) in human spermatozoa.

Progesterone induces a biphasic Ca(2+) influx and consequent acrosome reaction in human spermatozoa. We have used two procedures to vary the stimulus (dosage and prior receptor desensitization) to investigate the encoding of stimulus strength by intracellular [Ca(2+)] ([Ca(2+)](i)). Acrosome reaction and amplitude (but not kinetics) of the transient [Ca(2+)](i) response (population measurement) showed sigmoidal dose sensitivity over the range 0.3 nM-3 microM, saturating at approximately 300 nM (ED(50) approximately 30 nM). The amplitude of the sustained response saturated at 3 microM. Single-cell imaging showed that the amplitudes of both transient and sustained [Ca(2+)](i) responses were highly dose-dependent, but that their frequency of occurrence and kinetics were largely dose-independent. Fluorimetric measurements confirmed that progesterone-induced [Ca(2+)](i) influx was subject to desensitization, with second and subsequent applications of 3 microM progesterone being ineffective. However, sequential additions of 3 nM, 30 nM and 3 microM progesterone generated transient [Ca(2+)](i) responses at each concentration, the amplitude and duration of the response to 3 microM progesterone being reduced compared with non-pretreated cells. Single-cell imaging revealed that pretreatment had no effect on the proportion of responsive cells, but single-cell responses, similarly to population responses, were smaller and markedly reduced in duration, consistent with an effect of desensitization on a late component of the [Ca(2+)](i) transient. We conclude that the strength of the progesterone stimulus, when varied by dosage or by desensitization, is encoded by an analogue [Ca(2+)](i) signal. Dose dependency of the acrosome reaction is therefore determined not by the number of progesterone-responsive cells but by variation in the probability of exocytosis in a 'constant' responsive population.

Acrosome Reaction↗