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The afferent path of the milk-ejection reflex has been studied in the brain of the lactating guinea-pig in light pentobarbitone anaesthesia. Square-wave pulses were applied between an indifferent electrode in the scalp and a monopolar electrode inserted stereotaxically in the brain. The brain was transected at the mid-cerebellar level to eliminate activation of the sympathetico-adrenal system, and milk-ejection pressure was monitored to detect release of neurohypophysial hormone(s).

The afferent path of the reflex in the caudal midbrain was very compact and lay in the lateral tegmentum. More rostrally, milk-ejection responses were obtained from the tectum and mesencephalic central grey, but the major pathway remained in the lateral tegmentum and passed forward to lie ventromedial to the medial geniculate body, after which it divided into two components which we have termed the dorsal and ventral paths.

The dorsal path traversed dorsomedially across the brainstem to reach the parafascicular thalamic nucleus, the extreme rostral central grey and the periventricular region at the meso-diencephalic boundary, and then continued forward to reach the pituitary stalk and the medial and dorsal hypothalamus. The ventral path traversed ventromedially to enter the subthalamus and then the lateral hypothalamus, in which it passed both to the rostral basal diencephalon and to the pituitary stalk.

In the diencephalon, milk-ejection responses were obtained after stimulation of part of the ventral thalamus, the lateral, dorsal and anterior hypothalamic areas, the dorsomedial, ventromedial, arcuate, supraoptic and paraventricular nuclei, and the pituitary stalk.

It is suggested from these findings that in the guinea-pig the suckling stimulus ascends by the spinothalamic system, and continues rostrally to relay with the medial and ventral thalamus, the dorsal longitudinal fasciculus and the medial forebrain bundle. Other ascending pathways in the medial lemniscus and mammillary peduncle may also be involved, but appear to be of only minor significance.