314 EDWARD C. DAY 



The three following protocols will give the history of such an 

 operation with its subsequent effects both on the amputated 

 siphons and on the desiphonated body: 



Protocol 1. Ascidia mentula nos. 10, 11, and 12. April 17, 1913. 

 Narcotized all three animals with cocaine in sea-water and cut off the 

 tips of the siphons at XX' on the oral or incurrent siphon (distal to 

 oral tentacles) and at YY' on the aboral or excurrent siphon (fig. 1). 

 Returned animals to running sea-water. 



April 18. Amputated pieces A and C are sensitive to tactile stimu- 

 lation (B pieces not tested). 



April 19. A and C respond to tactile stimulation, but not to tapping 

 on the jar containing them. One of the bodies B is sensitive to the 

 tapping and responds by ejecting water from the siphons. Pharyn- 

 geal sacs have fallen away from the cut surfaces of the siphons. 



April 23. A and C pieces are dead. (The supply of running water 

 had got accidentally shut off.) The body pieces, B, are alive and 

 appear to be regenerating new tips. 



May 9. New siphons have been formed on all three animals. These 

 lie within the old cut tips and appear to have been developed from the 

 pharyngeal sac. Distinct lip-lobes and interlobular pigment spots are 

 visible on most of them: 7 to 8 lobes on IOC, 8 on 11 A, 6 on IIC, 5 

 on 12A; on lOA and 12C the siphons are too puckered up to make out 

 the number. 



Protocol 2. Ascidia m. no. 5. April 12, 1913. Cocainized the 

 animal (400 cc. sea-water -f 1 cc. 5 per cent cocain), and amputated 

 oral siphon below the circlet of oral tentacles at XX' (fig. 2). Pro- 

 cedure was as follows: 



5:10 P.M. Animal placed in cocain solution. 



6:10 P.M. Benumbed; operated on, rinsed in fresh sea-water and 

 returned to running sea-water. 



6:45 P.M. Excurrent siphon sensitive to tactile stimulation; ampu- 

 tated incurrent siphon A, unresponsive. 



April 15. A is responsive around marginal lobes; tentacles not. 

 Lobes pucker in in response to tactile stimulation and to rap on sub- 

 stratum. B is very sensitive both on cut surface and on excurrent 

 siphon. 



April 16. A and B respond to tap on jar. 



April 17. B responds to tap on jar; A, too, if tap is strong. 



April 19. B responds to tap on jar; A no longer. 



May 9. B has regenerated a new incurrent siphon with six small 

 lip-lobes and a new ring of pharyngeal tentacles. New siphon puckers 

 shut upon tapping the jar. 



Protocol 3. Ascidia mentula no. 17. May 10. Incurrent siphon 

 amputated to include the nerve ganglion. (Since the object of this 

 experiment was to study the effects of the operation on the heart beat, 

 no attention was paid to the amputated piece.) 



June 3. A new incurrent siphon and ganglion have regenerated; 

 six lobes to the siphon (fig, 3). 



